LINKAGE RELATIONS IN MALTING BARLEY

By

Arden D ex te r Day

AN ABSTRACT

S u b m itted to t h e S chool o f G rad uate S t u d i e s o f
M ichigan S t a t e C o lleg e of A g r i c u l t u r e and
A p p lie d S c ie n c e in p a r t i a l f u l f i l l m e n t
o f t h e r e q u ir e m e n ts f o r th e degree o f

DOCTOR OF PHILOSOPHY

D epartm ent o f Farm Crops

1954

Approved__

s .

""S '

A rd e n D e x te r Day

THESIS ABSTRACT

T h is r e s e a r c h problem was d e s ig n e d to s tu d y t h e
l i n k a g e r e l a t i o n s betw een d i a s t a s e a c t i v i t y , w hich i s
a m easure o f m a l t i n g q u a l i t y i n b a r l e y , and s p e c i f i c
v i s i b l e c h a r a c t e r i s t i c s known to be i n h e r i t e d on
c e r t a i n chromosome g ro u p s.

The f o llo w in g c r o s s e s

were made betw een s t a n d a r d b a r l e y v a r i e t i e s show­
in g t h e o p p o s it e e x p r e s s i o n o f a v i s i b l e c h a r a c t e r ­
i s t i c known t o be i n h e r i t e d on a p a r t i c u l a r chromo­
some g ro u p :

Chromosome group I ;
X

ALPHA

0 .A .C .2 1
( 6-row)

( 2-row)

Chromosome group I I :
X

DORSETT

0 .A .C .2 1
(W hite glumes)

(B lack glumes)

Chromosome group I I I ;
X

0 • A . C. 21

(Covered k e r n e l s )

-

1

-

C . I . 1370
(Naked k e r n e l s )

A rd en D e x te r Day

Chromosome group IV;
O .A.C.21

X

(Blue a le u r o n e )

MARS
(W hite a le u r o n e )

Chromosome group 7 :
O.A.C.21

X

(Rough awns)

MARS
(Smooth awns)

F iv e chromosome g ro u p s were r e p r e s e n t e d i n t h e
f i v e c r o s s e s , and O.A.C.21 was one of t h e p a r e n t s
o c c u r in g i n each c r o s s .

The p u re b r e e d i n g l i n e s

from th e Fg p o p u l a t i o n (F^ seed) in each c r o s s were
saved and a n a ly z e d f o r d i a s t a t i c power u s i n g a
m o d i f i c a t i o n o f th e A n d e rs o n -S a lla n s f e r r i c y a n i d e
m ethod.

P r o g e n ie s from tw e lv e s e l e c t i o n s from

ea ch o f th e two s ta n d a r d v a r i e t i e s used as p a r e n t s
in each c r o s s were a l s o c a r r i e d a lo n g i n th e same
f i e l d w ith t h e c r o s s e d m a t e r i a l and a n a ly z e d f o r
d i a s t a t i c power.

A f t e r th e ch em ical d e t e r m i n a t i o n s

were com pleted and t h e d i a s t a s e a c t i v i t y o f each
sample was c o n v e rte d i n t o d e g re e s L i n t n e r ( ° L ) , th e
two p a r e n t s o f each c r o s s and t h e two g ro u p s o f pure
b r e e d i n g progeny from each c r o s s were compared f o r
s i g n i f i c a n t d i f f e r e n c e s by means o f t h e t - t e s t .

- 2 -

A rd e n D e x te r Day

The r e s u l t s may be summarized as f o llo w s :
Chromosome group I :
The O.A.C.21 p a r e n t i s s i g n i f i c a n t l y h ig h e r i n
d i a s t a t i c power th a n t h e Alpha p a r e n t .

T here i s no

a p p a r e n t l i n k a g e between h ig h d i a s t a t i c power and
t h e 2-row c h a r a c t e r i s t i c .

T here i s an a p p a r e n t

p l e i o t r o p i c e f f e c t betw een h ig h d i a s t a t i c power
and th e 2-row c h a r a c t e r i s t i c .
Chromosome group I I :
The O.A.C.21 p a r e n t i s s i g n i f i c a n t l y h ig h e r in
d i a s t a t i c power th a n th e D o r s e tt p a r e n t .

T here i s

no s i g n i f i c a n t l i n k a g e betw een h ig h d i a s t a t i c power
and th e w h ite glume o r b la c k glume c h a r a c t e r i s t i c .
Chromosome group I I I :
The O.A.C.21 p a r e n t i s s i g n i f i c a n t l y h ig h e r in
d i a s t a t i c power t h a n t h e C . I . 1370 p a r e n t .

There i s

no s i g n i f i c a n t l i n k a g e betw een h ig h d i a s t a t i c power
and t h e co v e re d k e r n e l o r naked k e r n e l c h a r a c t e r i s t i c .
Chromosome group IV:
The O.A.C.21 p a r e n t i s s i g n i f i c a n t l y h i g h e r in
d i a s t a t i c power th a n th e Mars p a r e n t .

T here i s no

s i g n i f i c a n t l i n k a g e betw een h ig h d i a s t a t i c power and
t h e b lu e a l e u r o n e o r w h ite a le u r o n e c h a r a c t e r i s t i c .
However, t h e w h ite a le u r o n e group o f p u re b r e e d in g

- 3 -

A rd e n D e x te r Day

p ro g en y d id t e n d to ru n a l i t t l e h i g h e r i n d i a s t a t i c
power t h a n d id th e b l u e a le u r o n e g ro u p .
Chromosome group V:
The O.A.C.21 p a r e n t i s s i g n i f i c a n t l y h ig h e r in
d i a s t a t i c power th a n t h e Mars p a r e n t .

There i s an

a p p a r e n t l i n k a g e between h ig h d i a s t a t i c power and
th e rough awn c h a r a c t e r i s t i c .

- 4 -

LINKAGE RELATIONS IN MALTING BARLEY

By

Arden D e x te r Day

A THESIS

S u b m itte d to th e S cho ol o f G radu ate S tu d ie s o f
M ichigan S t a t e C o lle g e o f A g r i c u l t u r e and
A p plied S c ie n c e in p a r t i a l f u l f i l l m e n t
o f t h e r e q u ir e m e n ts f o r t h e d eg ree o f

DOCTOR OF PHILOSOPHY

D epartm ent o f Farm Crops

1954

ProQuest Number: 10008291

All rights reserved
INFORMATION TO ALL USERS
The quality of this reproduction is dependent upon the quality of the copy submitted.
In the unlikely event that the author did not send a complete manuscript
and there are missing pages, these will be noted. Also, if material had to be removed,
a note will indicate the deletion.

uest.
ProQuest 10008291
Published by ProQuest LLC (2016). Copyright of the Dissertation is held by the Author.
All rights reserved.
This work is protected against unauthorized copying under Title 17, United States Code
Microform Edition © ProQuest LLC.
ProQuest LLC.
789 East Eisenhower Parkway
P.O. Box 1346
Ann Arbor, Ml 48106-1346

- r sbi,
~ t> 3 - 1 3

ACKNOWLEDGSOTTS

T h is s tu d y was c a r r i e d o u t u n d er t h e g u id a n ce
o f Dr. E. E. Down and Dr. K. J . F r e y .

I ta k e t h i s

o p p o r t u n i t y t o e x p r e s s my s i n c e r e g r a t i t u d e to t h e s e
two men.

The w r i t e r i s p a r t i c u l a r l y in d e b te d t o Dr.

E. J . Benne f o r h i s s u p e r v i s i o n o f t h e ch e m ic al a n ­
a ly se s.

A p p r e c ia tio n i s ex ten d ed t o Dr. W. D. B aten

and P r o f e s s o r H. M. Brown f o r h e lp w ith th e s t a t i s ­
t i c a l a n a ly sis.

G -reatful acknowledgment i s g iv e n to

th e Midwest B a r le y Improvement A s s o c i a t i o n and th e
M ichigan B rew ers A s s o c i a t i o n f o r t h e i r c o o p e r a t i o n .
I a l s o e x p r e s s a s p e c i a l n o te o f g r a t i t u d e to my
w i f e , J e a n , f o r h e r c o n tin u e d a s s i s t a n c e and en­
couragement th r o u g h o u t t h e e n t i r e program .

To a l l

o t h e r s who c o n t r i b u t e d t o t h e s tu d y b u t go unmen­
t i o n e d , I am d e e p ly g r a t e f u l .

062370

VITA

Arden D e x te r Day
c a n d id a te f o r t h e d e g ree o f
D o ctor o f P h ilo s o p h y

F i n a l E x a m in atio n :
A g r i c u l t u r a l H a l l , M ichigan S t a t e C o lle g e , 1954.
D isse rta tio n :
L inkage R e l a t i o n s in M a ltin g B a r le y .
O u tlin e o f S t u d i e s :
M ajor F i e l d :
Minor F i e l d :

Farm Crops { P la n t B r e e d in g ) .
P l a n t P a th o lo g y and P l a n t P h y s io lo g y .

B i o g r a p h i c a l Ite m s :
B o m , March 16, 1922, West R u tla n d , Vermont.
U n d erg ra d u ate S tu d y , C o r n e ll U n i v e r s i t y , 1 9 4 6-5 0.
G raduate S tu d y , M ichigan S t a t e C o lle g e , 1 9 5 0-5 4.
E x p e r ie n c e :
F i e l d A s s i s t a n t , C o r n e ll U n i v e r s i t y , 1948-49.
F i e l d I n s p e c t o r , N.Y. S t a t e C e r t i f i e d Seed
Growers Coop., summer 1950.
G ra d u ate T each ing A s s i s t a n t , M ichigan S t a t e
C o l le g e , 1950 -54 .
G rad u ate R e se a rc h A s s i s t a n t , M ich ig an S t a t e
C o lle g e , summers 1951-53.
Member o f P i Mu E p s i l o n .

TABLE OF CONTENTS

Page
INTRODUCTION..........................................................................

1

REVIEW OF LITERATURE ......................................................

3

EXPERBIENT&L PROCEDURE..................................................

9

EXPERIMENTAL RESULTS............................................................ 20
..................................................21

Chromosome Group

I

Chromosome Group

I I ....................

Chromosome Group

I I I ..................................................26

Chromosome Group

I V ..................................................27

Chromosome Group

V

25

................................................. 28

DISCUSSION...................................................................................29
SUMMARY............................................................................................35
LITERATURE CITED ................................................................

39

1

INTRODUCTION

M a ltin g b a r l e y ( Hordeum v u lg a r e L .) i s t h e
p r i n c i p a l g r a i n u s e d i n t h e p r o d u c tio n o f m a lt w hich,
i n t u r n , i s th e b a s i c m a t e r i a l u se d i n t h e brew ing o f
b e e r and o t h e r a l c o h o l i c b e v e ra g e s .

M alt i s m a l t i n g

b a r l e y g r a i n which has b ee n s te e p e d in w a te r , germ­
i n a t e d u n d e r s p e c i f i c a l l y c o n t r o l l e d te m p e r a tu r e ,
m o i s t u r e , and tim e c o n d i t i o n s , and th e n d r i e d by a
p r o c e s s known a s k i l n i n g .
D uring t h e g e r m in a tio n o f b a r l e y g r a i n d e f i n i t e
p h y s i c a l and chem ical changes ta k e p la c e w i t h i n t h e
k e rn e l.

One o f th e m a jo r ch an g es i s an i n c r e a s e in

th e a c t i v i t y o f t h e enzyme d i a s t a s e w hich i s t h e
enzyme system t h a t changes s t a r c h i n t o s u g a r .

D ia sta se ,

commonly m easured in " d e g re e s L i n t n e r " ( ° L ) , i s made up
p r i m a r i l y o f two enzymes, a lp h a -a m y la se and b e t a - a m y la s e .
The amount o f d i a s t a s e i n m a lt i s c l o s e l y c o r r e l a t e d to
t h e amount o f s t a r c h t h a t can be d i g e s t e d w ith a g iv e n
amount o f m alt and th e r a t e a t which t h e d i g e s t i o n i s
a c c o m p lish e d .
I n t h i s r e s e a r c h problem an a tte m p t was made to
s tu d y th e l i n k a g e r e l a t i o n s betw een d i a s t a s e a c t i v i t y ,

2

which i s a m easure o f m a ltin g q u a l i t y in b a r l e y , and
sp e c ific v is ib le c h a ra c te ris tic s
on c e r t a i n chromosome g r o u p s .

known t o be i n h e r i t e d

3

HB7IBW OF LITERATURE

The e v o l u t i o n o f c u l t i v a t e d b a r l e y i s t i e d to the
p a s t b y t h r e a d s o f h i s t o r y as a n c ie n t a s th e o r i g i n o f
a g ric u ltu re i t s e l f .

The developm ent of t h i s crop

has

been molded i n th e hands o f many c u l t u r e s and h as been
accom panied by th e emergence o f a h o s t o f v a r i e t a l forms
i n many en v iro n m en ts th r o u g h o u t t h e w o rld .

B a r l e y has

alw ay s ta k e n an im p o r ta n t p la c e a lo n g t h e a g r i c u l t u r a l
fro n tie r.

The s ta te m e n t i s o f t e n made, and p r o b a b ly

w ith j u s t i f i c a t i o n , t h a t t h i s

c e r e a l h as

a w id er

e c o lo g ic ra n g e t h a n any o th e r

g r a i n crop

(2 5 ).

M a ltin g b a r l e y i s a s p e c i a l ty p e o f b a r l e y g r a i n
p o s s e s s i n g c e r t a i n p h y s i c a l and ch em ical q u a l i t i e s r e ­
q u ir e d by m a l t s t e r s and b rew ers ( 1 4 ) .

The p r i n c i p a l

g r a i n u se d i n t h e p r o d u c tio n o f m a l t , w hich i s t h e
b a s i c m a t e r i a l f o r brew ing b e e r , i s t h i s ty p e o f b a r l e y
(2 4 ).

There a r e s e v e r a l re a s o n s f o r t h e g r e a t p o p u l a r i t y

o f b a r l e y m a lt i n th e brew ing

in d u s t r y . F i r s t ofa l l ,

b a r l e y i s one o f th e h a r d i e s t

o f th e sm a ll g r a i n s .

It

can be more e a s i l y m a lte d f o r b rew in g p u r p o s e s th a n any
o t h e r c e r e a l , and t h e s o l u b l e s e x t r a c t e d from t h e m a lt
a r e o f a more d e s i r a b l e c h a r a c t e r th a n th o s e e x t r a c t e d

4

from o tiie r g r a i n s .

B a rle y m a lt c o n t r i b u t e s to sm oother

p e rfo rm a n c e i n th e brew house b y i t s q u ic k c o n v e rs io n
o f s t a r c h and i t s p r o d u c t i o n o f e a s i l y s t r a i n e d mash.
I n a d d i t i o n , b a r l e y m a lt h a s s u f f i c i e n t enzym atic
c a p a c i t y f o r t h e c o n v e rs io n o f th e i n s o l u b l e m a t e r i a l s ,
b o th in th e m a lt i t s e l f and i n t h e a d d i t i o n a l a d j u n c t s
added t o t h e m a lt, i n t o s o l u b l e s u b s t a n c e s .

The s u p e r i o r

v a lu e o f b a r l e y m a lt i n t h e b rew in g o f b e e r was re c o g n iz e d
e a r l y i n t h e 1 5 th c e n tu r y when t h e o r i g i n a l s e t t l e r s came
t o America ( 2 6 ) .
One o f th e p r i n c i p a l m e asu rin g s t i c k s u se d to d e te rm in e
m a l t i n g q u a l i t y i n b a r l e y i s d i a s t a s e a c t i v i t y ( 1 4 ,2 2 ) .
Hopkins and K rause (13) v e r i f y t h e im portance o f d i a s t a t i c
power a s a measure o f m a l t i n g q u a l i t y in b a r l e y .

D ia s ta s e

a c t i v i t y was one o f th e m easu res o f m a l t i n g q u a l i t y used
by L ejeu ne et_ a l .

(15) in t h e i r s tu d y o f methods o f b r e e d ­

in g new m a l t i n g b a r l e y v a r i e t i e s .

A fte r re a d in g th e

l i t e r a t u r e and v i s i t i n g m a lt h o u se s and b r e w e r i e s , i t
becomes v e ry a p p a r e n t t h a t d i a s t a s e a c t i v i t y i s one
c h a r a c t e r i s t i c o f m a lt t h a t d e s e r v e s u tm o st c o n s i d e r a t i o n
i n t h e m a l t i n g and b rew in g i n d u s t r i e s .
A f t e r t h e im p o rta n c e o f d i a s t a t i c power became f i r m l y
e s t a b l i s h e d , t h e n e x t problem was to f i n d an e f f i c i e n t and

5

a c c u r a t e method o f t e s t i n g t h e e n z y m atic a c t i v i t y o f
d i f f e r e n t sam ples o f b a r l e y g r a i n and m a l t .

I n th e

e a r l y 1920*s Rumsey (19) d e v is e d a lo n g , t e d i o u s
copper r e d u c t i o n method f o r m e a s u rin g t h e d i a s t a s e
a c t i v i t y o f wheat f l o u r u sed f o r b re a d m aking, b u t
t h i s method was n o t v e r y e f f i c i e n t .

L a te r B lis h e t

a l . (8) r e p o r t e d a p i c r i c a c i d c o l o r i m e t r i c te c h n iq u e
f o r e s t i m a t i n g d i a s t a t i c power a s b e in g p r e f e r a b l e to
any o f t h e v a r io u s cop per r e d u c t i o n m ethods from th e
s t a n d p o i n t o f con v en ien ce w ith o u t l o s s o f a c c u r a c y .
About 1923 Hagedorn and Je n sen (10) p ro p o sed a
f e r r i c y a n i d e m icro-m ethod f o r t h e e s t i m a t i o n o f b lo o d
su g ar.

Hanes (11) and o t h e r s found t h a t t h e H ag edo m -

J e n s e n f e r r i c y a n i d e method o f d e te r m in in g r e d u c in g
s u g a r s in a n im a l t i s s u e s and p r o d u c ts i s e q u a l l y w e l l
s u i t e d t o i n v e s t i g a t i o n s w ith p l a n t s and p l a n t p r o d u c t s .
Minor m o d i f i c a t i o n s and a d a p t a t i o n s can u s u a l l y be made
to s u i t s p e c i a l p u r p o s e s and r e q u ir e m e n ts .

In th e e a rly

1930*s B l is h and S a n d s te d t (7) m o d ifie d t h e H agedornJ e n s e n f e r r i c y a n i d e method of d e te r m in in g r e d u c i n g s u g a rs
(10) so t h a t i t c o u ld be used to e s t i m a t e d i a s t a s e a c t i v i t y
i n w heat f l o u r .

T h is method was b a s e d upon t h e r e d u c t i o n

o f f e r r i c y a n i d e to f e r r o c y a n i d e by r e d u c in g s u g a r s in

6

a lk a lin e so lu tio n .

A f t e r r e d u c t i o n was co m p leted , th e

amount o f s u g a r p ro du ced was d e term in ed hy a c i d i f y i n g
t h e s o l u t i o n w ith a c e t i c a c i d , a d d in g p o ta s s iu m io d id e
and s o l u b l e s t a r c h , and t i t r a t i n g w ith sodium t h i o s u l f a t e .
I n 1937, A nderson and S a l l a n s (3) p o in te d o u t t h a t
t h e o f f i c i a l method o f th e American S o c ie ty o f Brewing
C hem ists f o r th e d e t e r m in a t io n o f d i a s t a t i c power o f m a lt
( 2 ) , which was a l s o t e n t a t i v e l y ad o p ted by th e A s s o c i a t i o n
o f O f f i c i a l A g r i c u l t u r a l Chem ists ( 6 ) , and w hich was b e in g
s t u d i e d by th e A merican A s s o c i a t i o n o f C e re a l C h e m ists,
l e a v e s much to be d e s i r e d a s a r o u t i n e m ethod.

I t s un­

s u i t a b i l i t y was d e m o n stra te d by d a t a g iv e n by Coleman (9)
show ing t h a t se v e n te e n l a b o r a t o r i e s o b ta in e d r e s u l t s
v a r y i n g from 103°L. t o 158°L. f o r t h e same m a lt.

Anderson

and S a l l a n s (3) m o d ifie d t h e B l i s h - S a n d s t e d t f e r r i c y a n i d e
m ethod f o r d e te r m in in g t h e r e d u c in g power o f a d i g e s t e d
s t a r c h s o l u t i o n (7) t o p e r m it i t s u s e in th e d e t e r m in a t io n
o f t h e d i a s t a t i c power of m a lt i n d e g r e e s L i n t n e r ( ° L ) .
E x p e rim e n ta l d a t a i n d i c a t e d , t h a t u n d e r t h e c o n d i t i o n s o f
th e d e t e r m i n a t i o n , t h e f e r r i c y a n i d e method was s u p e r i o r to
any method y e t r e p o r t e d .
As a r e s u l t of i n v e s t i g a t i o n s made by S a l l a n s and
A nderson (2 0 , and e a r l i e r p a p e rs ) i t a p p e a r s t o be w e ll
e s t a b l i s h e d t h a t t h e d i a s t a s e a c t i v i t i e s , b o th f r e e and

7

t o t a l , o f b a r l e y g r a i n and m a lt a r e v a r i e t a l c h a r a c t e r ­
istic s.

D ata ac cu m u late d by A nderson e t a l .

H arris at a l .

(4) and

(12) who have i n v e s t i g a t e d d i a s t a s e

a c t i v i t y in b a r l e y and m a lt p r o v id e a d d i t i o n a l s u p p o r t
to t h i s h y p o t h e s i s .

S a l l a n s and A nderson (20) found

t h e p r o t e o l y t i c enzyme p a p a in t o be a good m a t e r i a l to
u s e to a c t i v a t e th e d i a s t a s e i n unm alted b a r l e y g r a i n ,
a f a c t l a t e r v e r i f i e d by o t h e r w o rk e rs ( 2 1 ) .

S a lla n s

and Anderson made d i a s t a s e a c t i v i t y d e te r m in a t io n s i n
144 s a u r i e s o f b a r l e y g r a i n and i n t h e m a lts made from
them u s i n g th e f e r r i c y a n i d e t e c h n i q u e .

The r e s u l t s ob­

t a i n e d were e x p re s s e d i n d e g r e e s L i n t n e r ( ° L ) .

These

b a r l e y sam ples r e p r e s e n t e d tw e lv e common v a r i e t i e s grown
i n Canada.

V a rie ta l d iffe re n c e s in d ia s ta s e a c tiv i ty

were d e m o n stra te d and t h e v a r i e t i e s t h a t were h ig h i n
b a r l e y d i a s t a s e a c t i v i t y were a l s o h ig h i n m a lt d i a s t a s e
a c tiv ity .

The t o t a l b a r l e y d i a s t a s e a c t i v i t y a p p e a re d

to b e a good i n d i c a t i o n o f what t h e t o t a l m alt d i a s t a s e
a c t i v i t y m ight be f o r any g iv e n v a r i e t y .
I n 1941, A nderson e t a l . (5) su g g e s te d t h a t i n th e
e a r l y s t a g e s o f a b a r l e y b r e e d in g program i t sh o u ld be
p o s s i b l e to t e s t th e b a r l e y g r a i n f o r d i a s t a s e a c t i v i t y
a f t e r a c t i v a t i n g i t w ith p a p a in , and e l im in a te th e m a l t i n g
t e s t , which r e q u i r e s c o n s id e r a b le tim e and s p e c i a l e q u ip -

e
m ent, w ith o u t any dan ger o f d i s c a r d i n g new l i n e s o f
b a r l e y w ith s u p e r i o r m a l t i n g q u a l i t y .

However, up

to t h i s tim e t h e r e was no e v id en ce t h a t h y b r id b a r l e y
s t r a i n s would r e a c t t h e same a s s ta n d a r d v a r i e t i e s t o
th e d ia s ta s e t e s t .
I n 1942, M e re d ith e t a l .

(16) r e p o r t e d t h a t b a r l e y

d i a s t a t i c pow er, a f t e r a c t i v a t i o n w ith p a p a in , g iv e s a
good i n d i c a t i o n o f m a lt d i a s t a t i c power f o r h y b r id l i n e s
of b a r l e y a s w e ll as f o r s ta n d a r d v a r i e t i e s .

I t was a l s o

s u g g e s te d , t h a t b y u s i n g t h i s t e s t f o r h i s h y b r id l i n e s
o f b a r l e y and com paring t h e s e v a lu e s w ith th o s e o f a good
s ta n d a r d m a l t i n g b a r l e y v a r i e t y , t h e p l a n t b r e e d e r may
o b t a i n u s e f u l in f o r m a tio n on h i s l i n e s much e a r l i e r th a n
i t c o u ld be o b ta in e d w ith s ta n d a r d m a ltin g t e s t s .

He

would th e n be a b l e to c o n c e n tr a t e on a re d u c e d p o p u l a t i o n
o f c o n s i d e r a b l e prom ise and o n ly t h e b e t t e r m a t e r i a l would
be i n c r e a s e d and s u b m itte d f o r m a l t i n g t e s t s .
R o b e rtso n e t a l .

(17, 18) have p u b lis h e d a summary o f

a l l o f t h e l i n k a g e s t u d i e s in b a r l e y which sh o u ld be v e ry
h e l p f u l in f o r m a ti o n f o r p l a n t b r e e d e r s who a r e i n t e r e s t e d
i n s tu d y i n g l i n k a g e r e l a t i o n s i n m a l t i n g b a r l e y .

9

EXPERIMENTAL PROCEDURE

I n th e summer o f 1950 th e f o llo w in g c r o s s e s w ere
made i n t h e f i e l d betw een s ta n d a r d b a r l e y v a r i e t i e s
showing t h e o p p o s it e e x p r e s s io n o f a v i s i b l e c h a r a c t e r i s t i c
known t o be i n h e r i t e d on a p a r t i c u l a r chromosome group:

Chromosome group I :
ALPHA.

O.A.C.21

( 2 - row)

( 6-row)

Chromosome group I I :
DORSETT

O.A.C.21

(B lack glumes)

(White glumes)

Chromosome group I I I :
O.A.C.21

X

C . I . 1370
(Naked k e r n e l s )

(Covered k e r n e l s )

Chromosome group IV:
O.A.C.21

MARS

(Blue a le u r o n e )

(White a le u r o n e )

Chromosome group Y:
O.A.C.21
(Rough awns)

X

MARS
(Smooth awns)

10

F iv e chromosome g roups were r e p r e s e n t e d i n th e f i v e
c r o s s e s , and O.A.C.21 was one o f t h e p a r e n t s o c c u r in g in
each c r o s s .

The F^ seed was sp ace p l a n t e d i n t h e g r e e n ­

house i n t h e w i n t e r o f 1 9 5 0-5 1.

The Fg seed was space

p l a n t e d i n th e f i e l d i n t h e summer o f 1951.

The Fg se ed

from each Fg p l a n t was h a r v e s t e d s e p a r a t e l y , and th e n
p l a n t e d in t h e f i e l d i n th e summer o f 1952 a s a s e p a r a t e
lin e .

S in c e t h i s t h e s i s problem was s e t up a s a l i n k a g e

s tu d y and d id n o t in v o lv e s e g r e g a t i o n , t h e s e g r e g a t i n g
p o p u l a t i o n was d i s c a r d e d .

Only t h e pure b r e e d in g l i n e s

from t h e Fg p o p u la tio n (F^ seed ) in each c r o s s were sav ed
and a n a ly z e d f o r d i a s t a t i c pow er.

P ro g e n ie s from tw elv e

s e l e c t i o n s from each o f th e two s ta n d a r d v a r i e t i e s o f
b a r l e y u se d as p a r e n t s i n each c r o s s were c a r r i e d a lo n g
i n t h e same f i e l d w ith th e c r o s s e d m a t e r i a l and a n a ly z e d
f o r d i a s t a t i c power.
The t a s k o f s e l e c t i n g a p p r o p r i a t e p a r e n t s f o r each
c r o s s , making t h e o r i g i n a l c r o s s e s , c a r r y i n g th e h y b r id
m a t e r i a l th r o u g h th e Fg g e n e r a t i o n , and i n c r e a s i n g each
l i n e u n t i l s u f f i c i e n t seed was o b ta in e d t o g r in d i n t o
f l o u r f o r chem ical a n a l y s i s , r e q u i r e d a p p r o x im a te ly
t h r e e y e a r s o f f i e l d and g reen h o u se work and in v o lv e d
over 1,000 l i n e s o f b a r l e y .
The ch em ical method u s e d to d e te rm in e t h e d i a s t a s e

11

a c t i v i t y o f t h e b a r l e y sam ples was a m o d i f i c a t i o n o f th e
A n d e r s o n - S a lla n s f e r r i c y a n i d e method ( 3 ) .

T h is m ethod,

w hich in v o lv e s t h e a c t i v a t i o n o f b a r l e y d i a s t a s e by t h e
p r o t e o l y t i c enzyme p a p a in , i s a r a t h e r lo n g p r o c e d u r e ,
t a k i n g about 36 h o u r s from t h e tim e t h e o r i g i n a l sample
i s w eighed o u t u n t i l a v a lu e f o r th e d i a s t a t i c power o f
th e b a rle y g ra in i s f i n a l l y o b ta in e d .

A fter t e s t i n g

d u p l i c a t e sam ples o f a s t a n d a r d v a r i e t y o f b a r l e y each
day f o r a b o u t t h r e e weeks and c a r e f u l l y o b s e r v in g t h e
r e s u l t s u n d e r d i f f e r e n t e n v iro n m e n ta l c o n d i t i o n s , i t
was found t h a t t h e te m p e r a tu r e and tim in g c o n d i t i o n s
a r e v e r y c r i t i c a l and must be c a r e f u l l y c o n t r o l l e d f o r
c o n siste n t r e s u lts .

T h is i s p a r t i c u l a r l y t r u e o f t h e

s t e p s i n v o l v i n g enzyme a c t i v i t y .

I t was f u r t h e r ob­

s e rv e d t h a t d u p l i c a t e sam ples o f t h e same b a r l e y ,
a n a ly z e d u n d e r c a r e f u l l y c o n t r o l l e d c o n d i t i o n s , sh o u ld
agree

w i t h i n 0 .3 m l. o f sodium t h i o s u l f a t e i n th e f i n a l

titra tio n .
A f t e r th e ch e m ic al p ro c e d u re was m a s te r e d , ea ch
b a r l e y sample was a n a ly z e d in d u p l i c a t e f o r d i a s t a t i c
pow er.

I f t h e two d u p l i c a t e s of each sample d id n o t

a g r e e w i t h i n 0 .3 m l. o f sodium t h i o s u l f a t e i n th e f i n a l
t i t r a t i o n , t h e y were d is c a r d e d and two new d u p l i c a t e s
were a n a ly z e d a g a in .

A s ta n d a r d v a r i e t y was t e s t e d a s

12

a check ea ch day and a s t a r c h s ta n d a r d was u sed as a
check on t h e s p e c i a l s t a r c h s o l u t i o n each week.

S ix

b a r l e y sam ples ( i n d u p l i c a t e ) , i n a d d i t i o n to th e ch e ck s,
were t h e maximum number t h a t cou ld be c a r r i e d e f f i c i e n t l y
a t one tim e .
The e n t i r e t a s k o f o b t a i n i n g t h e n e c e s s a r y equipm ent,
l e a r n i n g t h e chem ical p r o c e d u r e , and a n a ly z i n g th e b a r l e y
sam ples f o r d i a s t a t i c power to o k ab o u t one y e a r , r e q u i r e d
$700 w o rth o f s p e c i a l equipment and c h e m ic a ls , and in v o lv e d
ab o u t $3,500 w o rth o f ch e m ic al d e t e r m i n a t i o n s .
The r e a g e n t s used and t h e m ain s t e p s in v o lv e d i n t h e
c h e m ic a l p r o c e d u r e a r e g iv e n on t h e f o llo w in g p a g e s .

REAGENTS USED

S p e c i a l S t a r c h R eagent:
T h is s p e c i a l s t a r c h r e a g e n t sh o u ld be
p r e p a r e d a c c o r d in g t o th e s p e c i f i c a t i o n s
o f th e American S o c ie ty o f Brewing C hem ists

A c e ta te B u ffe r Reagent Used i n P r e p a r a t i o n o f
S p ecial S tarch :
D is s o lv e 68 grams of C.P. sodium a c e t a t e
i n 500 m l. o f N a c e t i c a c id and make up
to 1 l i t e r w ith d i s t i l l e d w a te r.

0.5N Sodium H ydroxide R eagent:
D is s o lv e 20 grams o f sodium h y d ro x id e in
1 l i t e r o f d i s t i l l e d w a te r .

A lk a lin e N/20 P e r r i c y a n i d e R eagent:
D is s o lv e 1 6 .5 grams o f p u re d r y p o ta s s iu m
f e r r i c y a n i d e and 22 grams o f anhydrous
sodium c a rb o n a te in 1 l i t e r o f d i s t i l l e d
w a te r .

Keep in a d ark g l a s s b o t t l e away

from l i g h t .

14

5.

A c e tic Acid R eagent:
T h is s o l u t i o n sh o u ld c o n t a in 200 m l. o f
g l a c i a l a c e t i c a c i d , 70 grams o f p o ta s s iu m
c h l o r i d e , and 20 grams o f z in c s u l f a t e p e r
lite r.

6.

P o ta s s iu m Io d id e R eagent:
To a 50$ s o l u t i o n o f p o ta s s iu m i o d i d e , add
one drop o f c o n c e n tr a t e d sodium h y d ro x id e
s o l u t i o n f o r each 100 m l. o f r e a g e n t t o
p r e v e n t d e t e r i o r a t i o n on s t a n d i n g .

The

r e a g e n t i s n o t f i t f o r use u n l e s s c o l o r l e s s .

7.

S o lu b le S ta r c h R eagent:
T h is r e a g e n t should c o n t a in 1$ s o lu b l e s t a r c h
i n a 30$ sodium c h l o r i d e s o l u t i o n .

P re p a r e

th e s o lu b l e s t a r c h s u s p e n s io n and p ou r s lo w ly
in to b o ilin g w a te r.

B o i l f o r two m in u t e s , add

th e sodium c h l o r i d e , and make t o volume.

The

r e a g e n t sh o u ld be t r a n s p a r e n t and c o l o r l e s s .

8.

N / 2 0 Sodium T h i o s u l f a t e R eag ent:

D is s o lv e 1 2 .4 1 grams o f sodium t h i o s u l f a t e in
1 l i t e r o f d i s t i l l e d w a te r .

S e l e c t o n ly c l e a r

c r y s t a l s from th e b e s t C .P . g ra d e .

15

Main S te p s I n v o lv e d i n t h e M o d ifie d A ndersonS a l l a n s F e r r i c y a n i d e Method o f D ete rm in in g
D i a s t a s e A c t i v i t y o f B a r l e y Grain

1.

Grind g r a i n t o p a s s th r o u g h a 20-mesh s c r e e n and
mix sample t h o r o u g h l y .

2.

C a r e f u l l y weigh 2 . 5 grams of b a r l e y f l o u r , 0 . 5 gram
o f p a p a i n , and p l a c e i n a 200 ml. v o l u m e t r i c s u g a r
f l a s k and mix t h o r o u g h l y .

3.

Add 50 ml. o f d i s t i l l e d w a t e r , mix, and p l a c e f l a s k
i n a w a t e r b a t h a t 20° C. f o r 2 1 .5 h o u r s .

4.

F i l t e r t h e sample t h r o u g h r a p i d f lo w in g f i l t e r p a p e r
and c o l l e c t t h e e x t r a c t .

5.

P i p e t t e 1 ml. o f b a r l e y e x t r a c t i n t o a 200 ml.
v o l u m e t r i c f l a s k c o n t a i n i n g 100 m l. o f s p e c i a l l y
b u f f e r e d s t a r c h s o l u t i o n (1) and mix t h o r o u g h l y .

6.

E x a c t l y 30 m in u t e s a f t e r add in g t h e b a r l e y e x t r a c t ,
add 10 ml. o f 0.5N sodium h y d ro x id e s o l u t i o n , mix,
make t o volume w i t h d i s t i l l e d w a t e r , and mix t h o r o u g h l y .

16

7,

P i p e t t e 5 ml. o f th e d i g e s t e d s t a r c h s o l u t i o n i n t o
a 100 m l , , w ide-mouth, hard g l a s s , E rlenm eyer f l a s k .

8,

Add e x a c t l y 10 ml. o f a l k a l i n e N/20 f e r r i c y a n i d e
s o l u t i o n t o t h e s t a r c h s o l u t i o n i n t h e Erlenmeyer
f l a s k and cover w i t h a sm a ll b e a k e r ,

9,

P l a c e t h e Erlenm eyer f l a s k i n a b a t h of v i g o r o u s l y
b o i l i n g w a te r f o r e x a c t l y 20 m i n u t e s .

10.

Im m ediate ly co o l t h e Erlenmeyer f l a s k and i t s c o n t e n t s
i n a b a t h o f cold w a t e r .

11.

Add 25 ml, o f a c e t i c a c i d r e a g e n t t o t h e Erlenmeyer
f l a s k and mix t h o r o u g h l y .

12.

Add 1 ml. o f 50$ KI, f o llo w ed by 2 ml. o f s o l u b l e
s t a r c h s o l u t i o n t o t h e f l a s k , and a g a i n mix t h o r o u g h l y .

13.

T i t r a t e w i t h N/20 sodium t h i o s u l f a t e t o t h e com plete
d i s a p p e a r a n c e o f t h e blue c o l o r and r e c o r d t h e number
o f ml. o f sodium t h i o s u l f a t e r e q u i r e d .

14.

Add 100 ml. o f t h e s p e c i a l , b u f f e r e d s t a r c h s o l u t i o n (1)
t o a n o t h e r 200 ml. v o l u m e t r i c f l a s k a s was done i n s t e p
number 5 b u t do n o t add any b a r l e y e x t r a c t t o t h i s f l a s k .
T h is i s a s t a r c h s t a n d a r d .

17

15.

C a rry t h e s t a r c h s t a n d a r d th r o u g h s t e p s 6 - 1 3
in clusiv e.

16.

S u b t r a c t t h e number o f ml. o f sodium t h i o s u l f a t e
re q u ire d to t i t r a t e the d ig e ste d s ta rc h s o lu tio n
( t h a t t o which t h e b a r l e y e x t r a c t was added) from
t h e number o f ml. o f sodium t h i o s u l f a t e r e q u i r e d
to t i t r a t e the s ta r c h standard.

The r e s u l t i n g

number i s t h e f e r r i e y a n i d e e q u i v a l e n t of t h e
d i a s t a t i c power o f t h e b a r l e y g r a i n .

17.

M u l t i p l y t h e sodium t h i o s u l f a t e e q u i v a l e n t by 36
t o c o n v e r t t h e r e s u l t s t o d e g r e e s L i n t n e r (°L)
which i s t h e s t a n d a r d way o f e x p r e s s i n g t h e
d i a s t a t i c power o f b a r l e y g r a i n and m a l t .

18

A f t e r t h e chemical d e t e r m i n a t i o n s were completed
an d t h e d i a s t a s e a c t i v i t y of each sample was c o n v e r t e d
i n t o d e g r e e s L i n t n e r ( ° L ) , t h e two p a r e n t s o f each c r o s s
and th e two g ro u p s o f p u r e b r e e d i n g progeny from each
c r o s s were compared f o r s i g n i f i c a n t d i f f e r e n c e s by means
o f the t - t e s t .
I n t h e com parison o f the two p a r e n t s o f ea ch c r o s s
t h e f o l l o w i n g symbols were used:
N

- The number o f s e l e c t i o n s from t h e
s t a n d a r d v a r i e t y used a s a p a r e n t .

M

- The mean.

S .D .

- The s t a n d a r d d e v i a t i o n , (sin gle determination)

t

- The t - v a l u e o f t h e t - t e s t .

I n th e com parison of t h e two groups of p u r e b r e e d i n g
progeny from each c r o s s the f o l l o w i n g symbols were u se d :
N

- The number o f l i n e s i n t h e p u re
b r e e d i n g group o f progeny.

M

- The mean.

S.D.

- The s t a n d a r d d e v i a t i o n , (sin gle determination)

t

- The t - v a l u e o f t h e t - t e s t .

Before r u n n i n g t h e t - t e s t ,

t h e means o f t h e naked

k e r n e l p a r e n t ( C . I . 1370) and t h e naked k e r n e l group o f
p u r e b r e e d i n g prog eny were a d j u s t e d f o r h u l l s u s i n g 13 %
a s t h e a v e r a g e p e r c e n t a g e of h u l l s on 6-row b a r l e y s ( 2 4 ) .

F ig u re 1,

T h is i s a p i c t u r e o f t h e s p e c i a l c o n s t a n t
t e m p e r a t u r e (20°G) w a t e r h a t h d e s i g n e d f o r u se
in the determ ination of d ia s ta s e a c t i v i t y in
b arley grain.

I t is constructed of s t a i n l e s s

s t e e l w i t h a s t a i n l e s s s t e e l s h e l f i n two
s e c tio n s (each s e c tio n 17” x 17” ) w ith 3 / 8 ”
d i a m e t e r p e r f o r a t i o n s to a l lo w w a te r c i r c u l a t i o n .
The b a t h i t s e l f i s 1 8 ” x 3 6 ” w i t h a d e p t h o f 1 2 ”
( in s id e dim ensions).

The w a l l s and bottom a r e

d o u b le w i t h a t h i c k n e s s o f § ” a n d a r e i n s u l a t e d
w i t h g l a s s w ool.

The s h e l v e s a r e a d j u s t i b l e f o r

h e i g h t by means o f s l o t t e d m e t a l l i c s t r i p s
a t t a c h e d t o t h e b a t h a t each c o r n e r o f ea ch
shelf.

The s h e l v e s r e s t on movable t r i a n g u l a r

m etal p ie c e s f i t t e d in to th e s l o t s .

This b a th

i s e q u ip p e d w i t h an a g i t a t o r , a s o l e n o i d v a l v e ,
a M e rc-to-M erc t h e r m o r e g u l a t o r , and a r e l a y
c o n t r o l box.

I t was p u r c h a s e d from George T.

Walker & Company, M i n n e a p o l i s , M i n n . , an d c o s t

19

F i g u r e 1.

20

EXPERBfENTAL RESULTS

21

Chromosome g r o u p I :

Cross:
ALPHA

X

( 2 - row)

0 .A .C .2 1
(6-row)

Parents:
0 .A .C .2 1

ALPHA.
N

-

12

N

-

M

- 183.06

M

- 215.13

S.D. -

21.79

S.D. -

12

2 4 .6 4

t - 3.37**

Pure B ree ding Progeny:
2-row Group

6-row Oroup

N

-

30

!T

-

M

- 211.09

i:

- 166.48

S.D. -

30.46

S.D. -

30

25.98

t - 6,10**

The 0.A .C .21 p a r e n t i s s i g n i f i c a n t l y h i g h e r i n
d i a s t a t i c power t h a n t h e Alpha p a r e n t .

T here i s no

a p p a r e n t l i n k a g e between h ig h d i a s t a t i c power and t h e
2-row c h a r a c t e r i s t i c .

T here i s an a p p a r e n t p l e i o t r o p i c

e f f e c t between h ig h d i a s t a t i c power and t h e 2-row
ch aracteristic.

**!<£ l e v e l o f s i g n i f i c a n c e

22

L et us s e t up a sim ple i l l u s t r a t i o n t o show t h a t
t h e r e i s no a p p a r e n t l i n k a g e between h i g h d i a s t a t i c
power and th e 2-row c h a r a c t e r i s t i c .

I f we assume, t h a t

head ty p e i s c o n t r o l l e d by 1 gene (Y a 2-row; v = 6 -row ),
t h a t d i a s t a t i c power i s c o n t r o l l e d by a n o t h e r s i n g l e gene
(A a h ig h ; a a l o w ) , t h a t t h e r e i s a p e r f e c t l i n k a g e of
t h e s e 2 genes on t h e same chromosome, and t h a t we c r o s s a
2-row, low d i a s t a s e p a r e n t (Alpha) w i t h a 6-row, h ig h
d i a s t a s e p aren t (0.A .C .21), then th e follow ing s i t u a t i o n
sh o u ld e x i s t :

V - a
V - a
Gametes

v - A
v - A

T

v - A

V - a

?1

Y
v

-a
-A

,Y

-a -

Y - a,

Y - a
Y - a

(2-row , low d i a s t a s e
p u r e b r e e d i n g progeny)

Y - a
v - A

,v - A
,Y - a

s

Y - a
v - A

,v - A

=

v A ( 6 - row, h i g h d i a s t a s e
v - A
p u r e b r e e d i n g progeny)

v - A,

From t h e p r e c e d i n g i l l u s t r a t i o n , i t i s c l e a r l y e v i d e n t ,
i f d i a s t a s e a c t i v i t y were l i n k e d w i t h 2-row head t y p e , t h e n
th e 2-row group o f p u r e b r e e d i n g progeny should have t h e lo w er
mean, which i s n o t t h e c a s e .

There i s no a p p a r e n t l i n k a g e

between h ig h d i a s t a s e a c t i v i t y and th e 2-row c h a r a c t e r i s t i c .

23

Let u s s e t up a sim p le i l l u s t r a t i o n t o show t h a t
t h e r e i s an a p p a r e n t p l e i o t r o p i c e f f e c t between d i a s t a t i c
power and th e 2-row and 6-row c h a r a c t e r i s t i c .

I f we

assume, t h a t head t y p e i s c o n t r o l l e d by 1 gene (V = 2-row;
v * 6 - r o w ) , t h a t d i a s t a t i c power i s c o n t r o l l e d by a n o t h e r
s i n g l e gene ( a * h i g h ; a = lo w ), t h a t t h e s e 2 genes a r e
i n d e p e n d e n t l y i n h e r i t e d on d i f f e r e n t chromosomes, and
t h a t we c r o s s a 2-row , low d i a s t a s e p a r e n t (Alpha) w ith
a 6-row, h ig h d i a s t a s e p a r e n t ( 0 . A . C . 2 1 ) , th e n t h e f o l l o w ­
i n g s i t u a t i o n should e x i s t ;

Waa

1

Gametes

X

vvAA
vA

Va
VvAa
WAA 1 )
WAa 2 )
Waa 1 )

Pure b r e e d i n g 2-row pro geny saved.

VvAA 2 )
VvAa 4 )
Waa 2 )

S e g r e g a t i n g 2-row and 6-row
p ro g en y d i s c a r d e d .

vvAA 1 )
vvAa 2 )
waa 1 )

P u re b r e e d i n g 6-row progeny saved.

From t h e p r e c e d i n g i l l u s t r a t i o n , i t i s e v i d e n t t h a t
t h e p u r e b r e e d i n g 2-row group o f progeny has more genes

24

f o r h i g h d i a s t a t i c power t h a n t h e 2-row p a r e n t ( A l p h a ) .
T h e r e f o r e , t h e 2-row group o f p ro g e n y would be e x p e c te d
to have a h i g h e r d i a s t a t i c power t h a n t h e Alpha p a r e n t .
L i k e w i s e , s i n c e t h e p ure b r e e d i n g 6-row group o f progeny
h a s fewer g ene s f o r h i g h d i a s t a t i c power t h a n t h e O.A.C.
21 p a r e n t , i t would be e x p e c t e d t o have a lower d i a s t a t i c
power t h a n O.A.C.21.

Up t o t h i s p o i n t , d i a s t a t i c power

seems t o be c o n t r o l l e d by two i n d e p e n d e n t l y i n h e r i t e d
factors.

However, i f one o b s e rv e s t h a t th e 6-row group

o f pro geny has a low er mean t h a n even t h e low er d i a s t a s e
p a r e n t ( A l p h a ) , i t a p p e a r s t h a t t h e 2-row gene i s a p p a r e n t l y
g i v i n g an a d d i t i o n a l i n c r e a s e t o t h e d i a s t a t i c power.

It

i s a l s o i n t e r e s t i n g t o n o t e t h a t t h e 6-row gene seems t o
have a g r e a t e r d e p r e s s i n g e f f e c t upon t h e d i a s t a t i c power
th a n the 2-row gene h a s an i n c r e a s i n g e f f e c t upon t h e
d i a s t a t i c power.

There i s an a p p a r e n t p l e i o t r o p i c

e f f e c t between d i a s t a t i c power and t h e 2-row and 6-row
ch aracteristic.

25

Chromosome g r o u p I I ;

Cross:
DORSETT

X

(Black glumes)

O .A .C .21
(White glumes)

P arents:
DORSETT

O .A.C.21

N

-

12

N

-

M

- 163.89

M

- 215.13

S.D. -

19.82

S.D. -

12

24.64

t - 5.61**

P u r e B r e e d in g P rogeny:
B lack Glume Group

White Glume G-roup

N

-

30

N

-

M

- 163.42

M

- 169.36

S.D. -

23.4 2

S.D. -

30

30.26

t - 0 .8 5

The O .A .C .21 p a r e n t i s s i g n i f i c a n t l y h i g h e r i n
d i a s t a t i c power t h a n t h e D o r s e t t p a r e n t .

T here i s no

s i g n i f i c a n t l i n k a g e between h i g h d i a s t a t i c power and
t h e w h i t e glume o r b l a c k glume c h a r a c t e r i s t i c .

* * 1 fo l e v e l o f s i g n i f i c a n c e

26

Chromosome g ro u p I I I :

Cro s s :
O .A .C .21

X

(Covered k e r n e l s )

C . I . 1370
(Naked k e r n e l s )

Parents:
C . I . 1370

0 . A.C.21
N

-

12

N

-

M

- 215.13

M

- 186.20

S.D. -

2 4 .6 4

S.D. -

12

16.00

t - 3.41**

P u r e B r e e d in g P rogeny:
COVERED KERNEL CROUP

NAKED KERNEL GROUP

N

-

N

-

M

- 145.73

M

- 150.71

S.D. -

5

13.45

S.D. -

5

14.90

t - 0 .5 5

The O .A .C.21 p a r e n t i s s i g n i f i c a n t l y h i g h e r i n
d i a s t a t i c power t h a n t h e C . I . 1370 p a r e n t .

There i s

no s i g n i f i c a n t l i n k a g e between h i g h d i a s t a t i c power and
t h e covered k e r n e l o r naked k e r n e l c h a r a c t e r i s t i c .

**1^ l e v e l o f s ig n if ic a n c e

27

Chromosome g r o u p IV :

C ross:
O.A.C. 21

X

(Blue a l e u r o n e )

MARS
(White a l e u r o n e )

P arents:
O.A.C.21

MARS

N

-

12

N

-

M

- 215.13

M

- 141.15

S.D. -

24.64

S.D. -

12

16.80

t - 8,58**

P u re B r e e d in g P rogeny:
BLUB ALEURONE G30TJP

WHITE ALEURONE GROUP

N

-

33

N

-

M

- 143.17

M

- 151.32

S.D. -

22.06

S.D. -

30

2 4 .7 5

t - 1.37

The O.A.C.21 p a r e n t i s s i g n i f i c a n t l y h i g h e r i n
d i a s t a t i c power t h a n t h e Mars p a r e n t .

T here i s no

s i g n i f i c a n t l i n k a g e between h ig h d i a s t a t i c power and
the b lu e aleurone or white aleurone c h a r a c t e r i s t i c .
However, t h e w h i t e a l e u r o n e group d i d t e n d t o run a
l i t t l e h i g h e r i n d i a s t a t i c power t h a n d id t h e b l u e
a l e u r o n e g ro u p .
* * 1 '% l e v e l o f s i g n i f i c a n c e

28

Chromosome g r o u p V:

Cross:
O.A.C. 21

X

(Rough awns)

MARS
(Smooth awns)

P arents:
O.A .C.21

MARS

N

-

12

N

-

M

- 215.13

M

- 141.15

S.D. -

24.64

S.D. -

12

16.80

t - 8.58**

Pure B r e e d in g P rogeny:
ROUGH AWN G-ROUP

SMOOTH AWN GROUP

N

-

32

N

-

M

- 151.29

M

- 128.44

S.D. -

3 0 .7 1

S.D. -

31

27 .4 6

t - 3.12**

The O.A.C.21 p a r e n t i s s i g n i f i c a n t l y h i g h e r i n
d i a s t a t i c power t h a n t h e Mars p a r e n t .

There i s an

a p p a r e n t l i n k a g e between h i g h d i a s t a t i c power and t h e
rough awn c h a r a c t e r i s t i c .

* * 1 fa l e v e l o f s i g n i f i c a n c e

29

DISCUSSION

I t i s e v id e n t, a f t e r observing the experim ental
r e s u lts , that d iastase a c tiv ity is d e fin ite ly a v a r ie ta l
ch aracteristic.

The two p a r e n t a l v a r i e t i e s i n v o lv e d i n

e v e r y c r o s s d i f f e r e d s i g n i f i c a n t l y from each o t h e r i n
diastase a c tiv ity .
linkage r e la tio n s h ip

There a l s o a p p e a r s t o be d e f i n i t e
between c e r t a i n v i s i b l e c h a r a c t e r ­

i s t i c s and h i g h d i a s t a t i c power, which i s a measure of
m alting q u a lity in b arley .
The v i s i b l e c h a r a c t e r i s t i c u se d i n t h e s tu d y o f
chromosome I was 2-row v s . 6-row head t y p e .

The two

s t a n d a r d v a r i e t i e s used i n t h i s f i r s t c r o s s were Alpha
and O .A .C.21.

The O.A.C.21 p a r e n t , a 6-row v a r i e t y ,

was s i g n i f i c a n t l y h i g h e r i n d i a s t a s e a c t i v i t y th a n Alpha.
Of t h e p u re b r e e d i n g progeny from t h i s c r o s s , t h e 2-row
group was s i g n i f i c a n t l y h i g h e r in d i a s t a t i c power t h a n
r

t h e 6-row g ro u p , which i n d i c a t e s an a p p a r e n t p l e i o t r o p i c
e f f e c t between h i g h d i a s t a s e a c t i v i t y and 2-row head
type,

p l e i o t r o p i s m i s t h e c o n t r o l l i n g of more t h a n one

c h a r a c t e r by a s i n g l e gene.

There i s no a p p a r e n t l i n k a g e

between h i g h d i a s t a s e a c t i v i t y and t h e 2-row c h a r a c t e r i s t i c .
I f h ig h d i a s t a s e a c t i v i t y and 2-row head ty p e were l i n k e d ,

30

t h e 2-row group o f pure b r e e d i n g progeny should show th e
lo w e s t mean b e c a u se t h i s i s t h e c o n d i t i o n t h a t e x i s t s i n
th e p a re n ts .
Cross number two in v o lv e d D o r s e t t and O.A .C.21, two
s t a n d a r d v a r i e t i e s o f b a r l e y showing a d i f f e r e n c e in
glume c o l o r .

D o r s e t t has b l a c k glumes w h i l e O .A .C .21

h a s w h ite glumes.

T h is v i s i b l e c h a r a c t e r i s t i c i s i n ­

h e r i t e d on chromosome I I .

The O.A.C.21 p a r e n t was

s i g n i f i c a n t l y h i g h e r i n d i a s t a t i c power th a n D o r s e t t .
Of t h e pure b r e e d i n g progeny from t h i s c r o s s , t h e r e
was no s i g n i f i c a n t d i f f e r e n c e i n d i a s t a t i c power
between t h e b l a c k glume group and t h e w h ite glume
group.

This i n d i c a t e s t h a t t h e r e i s no a p p a r e n t

l i n k a g e between h i g h d i a s t a t i c power and t h e w h ite
o r b l a c k glume c h a r a c t e r i s t i c .
O .A .C .21 and C . I . 1370 were t h e two s t a n d a r d
v a r i e t i e s use d as p a r e n t s i n t h e s t u d y o f chromosome I I I .
T h is t h i r d c r o s s in v o lv e d covered k e r n e l s v s . naked k e r n e l s .
O .A .C .21 i s a cove re d k e r n e l v a r i e t y , w h i l e C . I . 1370 has
naked k e r n e l s .

The O .A .C.21 p a r e n t was s i g n i f i c a n t l y

h i g h e r i n d i a s t a t i c power th a n t h e C . I . 1370 p a r e n t .

Of

t h e p u r e b r e e d i n g progeny from t h i s c r o s s , t h e r e was no
s i g n i f i c a n t d i f f e r e n c e i n d i a s t a t i c power between t h e
c o v e re d k e r n e l group and t h e naked k e r n e l group.

There­

f o r e , t h e r e i s no a p p a r e n t l i n k a g e between h i g h d i a s t a s e
a c t i v i t y and t h e covered o r naked k e r n e l c h a r a c t e r i s t i c .

31

Cross number f o u r in v o l v e d O.A.C.21 and Mars, two
s t a n d a r d v a r i e t i e s o f b a r l e y showing a d i f f e r e n c e i n
aleurone c o lo r.

O .A .C .21 has b lu e a l e u r o n e w h ile Mars

h as w h ite a l e u r o n e .

T h is v i s i b l e c h a r a c t e r i s t i c i s

i n h e r i t e d on chromosome IV.

The O.A.C.21 p a r e n t was

s i g n i f i c a n t l y h i g h e r i n d i a s t a t i c power th a n Mars,
The two g ro u p s of pure b r e e d i n g progeny from t h i s
c r o s s showed no s i g n i f i c a n t d i f f e r e n c e i n enzymatic
a c t i v i t y , a l t h o u g h t h e w h i t e a l e u r o n e group d i d show
a h i g h e r d i a s t a t i c power t h a n t h e b lu e a l e u r o n e group.
T h i s t e n d e n c y i s a l s o i n t e r e s t i n g b e c a u s e t h e w h ite
a l e u r o n e b a r l e y s seem t o be p r e f e r r e d f o r m a l t i n g
purposes.
The v i s i b l e c h a r a c t e r i s t i c use d i n t h e s t u d y o f
chromosome V was rough awns v s . smooth awns.

The two

s t a n d a r d v a r i e t i e s used i n t h i s case were O .A .C .21 and
Mars.

O.A.C.21 i s a rough awn v a r i e t y and Mars has

smooth awns.

The O.A .C.21 p a r e n t was s i g n i f i c a n t l y

h i g h e r i n d i a s t a t i c power th a n th e Mars p a r e n t .

Of

t h e p u r e b r e e d i n g progeny from t h i s c r o s s , t h e rough
awn group was s i g n i f i c a n t l y h i g h e r in d i a s t a t i c power
t h a n t h e smooth awn g ro up, which i n d i c a t e s an a p p a r e n t
l i n k a g e between h ig h d i a s t a s e a c t i v i t y and th e rough
awn c h a r a c t e r i s t i c .

T h is r e l a t i o n s h i p sh o u ld be h e l p f u l

32

i n f o r m a t i o n t o b a r l e y b r e e d e r s who a r e i n t e r e s t e d i n
s e le c tin g su p e rio r m alting b a rle y l i n e s in th e e a rly
s t a g e s o f t h e i r b r e e d i n g program.

For example, i f a

b a r l e y b r e e d e r should make a c r o s s between a rough awn,
h i g h d i a s t a t i c power v a r i e t y such a s M anchuria, K in d r e d ,
o r O d e r b ru c k e r and a smooth awn, low d i a s t a t i c power
v a r i e t y s u c h a s W isco nsin 38, Bay, o r Moore and i f t h e r e
i s a r a t h e r t i g h t l i n k a g e between h ig h d i a s t a t i c power
and rough awns, a s t h e d a t a i n t h i s t h e s i s i n d i c a t e ,
t h e n t h e b a r l e y b r e e d e r should be a b l e t o go i n t o h i s
b r e e d i n g n u r s e r y , s e l e c t th e most p r o m is in g l o o k i n g
l i n e s showing t h e r ough awn c h a r a c t e r i s t i c , and be
r e a s o n a b l y s u r e t h a t he i s a l s o s e l e c t i n g f o r h ig h
d iastase a c tiv ity .

On t h e o t h e r hand, i f a b a r l e y

b r e e d e r sh o u ld make a c r o s s between a rough awn, low
d i a s t a t i c power v a r i e t y such as T r e b i and a smooth awn,
h i g h d i a s t a t i c power v a r i e t y such as Montcalm, t h e n t h e
smooth awn c h a r a c t e r i s t i c should be used t o s e l e c t l i n e s
h a v i n g h i g h d i a s t a t i c power.

Therefore, i f a b arle y

b r e e d e r d e s i r e s to use t h i s a p p a r e n t l i n k a g e i n making
s e l e c t i o n s from h i s b r e e d i n g n u r s e r y , he must f i r s t
t e s t t h e p a r e n t s o f each c r o s s f o r d i a s t a t i c power so
he w i l l know which p a r e n t has t h e h i g h e s t d i a s t a t i c
power and which awn ty p e (ro u g h o r smooth) t o s e l e c t

33

f o r t o g iv e him h ig h d i a s t a t i c power i n t h e s e l e c t e d
p ro g e n y .
As was n o t e d by S a l l a n s and Anderson (20) i n
r e p o r t i n g t h e i r s t u d i e s on th e r e l a t i o n s h i p between
b a r l e y d i a s t a t i c power, a f t e r a c t i v a t i o n by p a p a i n ,
and m a lt d i a s t a t i c power, t h e r e i s much t o be g a in e d
i n a b a r l e y b r e e d i n g program , i f l i n e s can be s e l e c t e d
a t an e a r l y s t a g e on th e b a s i s of enzyme a c t i v i t y .

The

d e t e r m i n a t i o n o f a c t i v a t e d b a r l e y d i a s t a t i c power may
be made on a sample of 2 .5 grams a s compared t o 550 grams
n eeded f o r a m a l t i n g t e s t ( 1 6 ) .

I t i s evident th a t

samples can be t e s t e d f o r b a r l e y d i a s t a t i c power much
e a r l i e r and more c o n v e n i e n t l y t h a n th e y can be s u b j e c t e d
to m alting t e s t s .
The f e r r i c y a n i d e method of d e t e r m in in g t h e d i a s t a t i c
power o f b a r l e y g r a i n u se d i n t h i s problem proved t o be a
s a t i s f a c t o r y method f o r d e t e r m i n i n g t h e d i a s t a s e a c t i v i t y
o f m a l t i n g b a r l e y l i n e s and v a r i e t i e s .

S in ce b a r l e y

d i a s t a t i c power i s p o s i t i v e l y c o r r e l a t e d w i t h m a lt
d i a s t a t i c power i t can be used t o p r e d i c t t h e m a lt
d i a s t a s e a c t i v i t y of h y b r i d b a r l e y l i n e s .

In ad d itio n ,

s i n c e t h e f e r r i c y a n i d e t e s t r e q u i r e s o n l y a 2 .5 gram
sample o f ground b a r l e y , i t can be run v e r y e a r l y i n
t h e b r e e d i n g program, which would e n a b le t h e b a r l e y

34

b r e e d e r t o c o n c e n t r a t e on a reduced p o p u l a t i o n o f
c o n s i d e r a b l e p ro m ise .

He would t h e n i n c r e a s e and

submit o n ly h i s b e t t e r m a t e r i a l f o r m a l t i n g t e s t s .
Now t h a t t h e im p o r t a n t f a c t o r s a f f e c t i n g m a l t i n g
q u a l i t y i n b a r l e y have been d eterm in ed ( d i a s t a s e
a c t i v i t y , malt- e x t r a c t p e r c e n t a g e , p r o t e i n p e r c e n t a g e ,
e t c . ) and methods o f p r e d i c t i n g t h e s e f a c t o r s i n e a r l y
g e n e r a t i o n s have been d e v i s e d , t h e n e x t b i g s t e p i n
m a l t i n g b a r l e y r e s e a r c h sh o u ld be s t u d i e s to de te rm in e
w h e th e r o r n o t t h e r e a r e a d d i t i o n a l l i n k a g e r e l a t i o n ­
s h i p s between v i s i b l e c h a r a c t e r i s t i c s t h a t t h e b a r l e y
b r e e d e r can o b se rv e i n t h e f i e l d and t h e i m p o r ta n t
f a c to r s a f f e c t i n g m alting q u a l ity .

A dditional

s t u d i e s , such a s th e one r e p o r t e d i n t h i s t h e s i s ,
s h o u ld be c o n t in u e d and expanded so t h a t f u t u r e
b a r l e y b r e e d e r s w i l l know w h eth e r or n o t i t i s
p o s s i b l e f o r them t o s e l e c t f o r c e r t a i n v i s i b l e
c h a r a c te r i s tic s in t h e i r b arley breeding n u rseries
and be a s s u r e d t h a t t h e y a r e a l s o s e l e c t i n g f o r
m a l t i n g q u a l i t y a t t h e same tim e.

35

SUMMARY

T h is r e s e a r c h problem was d e s ig n e d t o s t u d y t h e
l i n k a g e r e l a t i o n s between d i a s t a s e a c t i v i t y , which i s
a measure o f m a l t i n g q u a l i t y i n b a r l e y , and s p e c i f i c
v i s i b l e c h a r a c t e r i s t i c s known t o be i n h e r i t e d on
c e r t a i n chromosome g r o u p s .

The f o l l o w i n g c r o s s e s

were made between s t a n d a r d b a r l e y v a r i e t i e s show­
in g t h e o p p o s i t e e x p r e s s i o n o f a v i s i b l e c h a r a c t e r ­
i s t i c known t o be i n h e r i t e d on a p a r t i c u l a r chromo­
some group:

Chromosome group I :
ALPHA

X

O.A.C.21
( 6 - row)

( 2 - row)

Chromosome group I I :
DORSETT

X

O .A .C.21
(White glumes)

( B la ck glumes)

Chromosome group I I I : .
O.A.C.21
(Covered k e r n e l s )

X

C . I . 1370
(Naked k e r n e l s )

36

Chromosome group IV:
O .A.C.SI

X

(Blue a l e u r o n e )

MARS
(White a l e u r o n e )

Chromosome group V:
O .A.C.21
(Rough awns)

X

MARS
(Smooth awns)

F iv e chromosome groups were r e p r e s e n t e d i n th e
f i v e c r o s s e s , and O.A.C.21 was one o f t h e p a r e n t s
o c c u r i n g i n each c r o s s .

The pure b r e e d i n g l i n e s

from t h e Fg p o p u l a t i o n (F4 seed) i n each c r o s s were
saved and a n a ly z e d f o r d i a s t a t i c power u s i n g a
m o d i f i c a t i o n o f t h e A n d e r s o n - S a lla n s f e r r i c y a n i d e
method ( 3 ) .

P r o g e n i e s from tw elve s e l e c t i o n s from

each o f t h e two s t a n d a r d v a r i e t i e s use d a s p a r e n t s
i n each c r o s s were a l s o c a r r i e d a l o n g i n t h e same
f i e l d w i t h t h e c r o s s e d m a t e r i a l and a n a ly z e d f o r
d i a s t a t i c power.

A f t e r t h e chemical d e t e r m i n a t i o n s

were com pleted and t h e d i a s t a s e a c t i v i t y o f each
sample was c o n v e r t e d i n t o d e g r e e s L i n t n e r ( ° L ) , t h e
two p a r e n t s o f e a c h c r o s s and t h e two groups of pure
b r e e d i n g progeny from each c r o s s were compared f o r
s i g n i f i c a n t d i f f e r e n c e s by means of t h e t - t e s t .

37

The r e s u l t s may be summarized as f o l l o w s :
Chromosome group I :
The O.A .C.El p a r e n t i s s i g n i f i c a n t l y h i g h e r i n
d i a s t a t i c power t h a n t h e Alpha p a r e n t .

There i s no

a p p a r e n t l i n k a g e between h i g h d i a s t a t i c power and
t h e 2-row c h a r a c t e r i s t i c .

There i s an a p p a r e n t

p l e i o t r o p i c e f f e c t between high d i a s t a t i c power
and t h e 2-row c h a r a c t e r i s t i c .
Chromosome group I I ;
The O.A.C.21 p a r e n t i s s i g n i f i c a n t l y h i g h e r in
d i a s t a t i c power t h a n t h e D o r s e t t p a r e n t .

There i s

no s i g n i f i c a n t l i n k a g e between h ig h d i a s t a t i c power
and t h e w h i t e glume o r b l a c k glume c h a r a c t e r i s t i c .
Chromosome group I I I :
The O .A .C .21 p a r e n t i s s i g n i f i c a n t l y h i g h e r in
d i a s t a t i c power t h a n t h e C . I . 1370 p a r e n t .

There i s

no s i g n i f i c a n t l i n k a g e between h ig h d i a s t a t i c power
and t h e covered k e r n e l or naked k e r n e l c h a r a c t e r i s t i c .
Chromosome group 1 7 :
The O.A .C.21 p a r e n t i s s i g n i f i c a n t l y h i g h e r i n
d i a s t a t i c power t h a n t h e Mars p a r e n t .

There i s no

s i g n i f i c a n t l i n k a g e between h i g h d i a s t a t i c power and
t h e b l u e a l e u r o n e or w h ite a l e u r o n e c h a r a c t e r i s t i c .
However, th e w h ite a l e u r o n e group o f pure b r e e d i n g

38

p r o g e n y d i d t e n d t o run a l i t t l e h i g h e r i n d i a s t a t i c
power t h a n d i d t h e b lu e a l e u r o n e group.
Chromosome group V;
The 0.A .C .2 1 p a r e n t i s s i g n i f i c a n t l y h i g h e r i n
d i a s t a t i c power t h a n th e Mars p a r e n t .

There i s an

a p p a r e n t l i n k a g e between h i g h d i a s t a t i c power and
t h e rough awn c h a r a c t e r i s t i c .

39

LITERATURE CITED

1.

American S o c i e t y of Brewing C hemists.
o f a n a l y s i s . P p . 99-105. 1949.

Methods

2.

American S o c i e t y o f Brewing Chemists.
m ethods. 1936.

O fficial

3.

Anderson, J . A . , and H. R. S a l l a n s . D e te r m i n a t i o n
o f t h e d i a s t a t i c power of m a lt i n d e g r e e s L i n t n e r
by means o f a f e r r i c y a n i d e r e a g e n t . Can. J .
R e s e a rc h , C, 15; 7 0-77. 1937.

4.

A nderson, J . A ., H. R. S a l l a n s ,
and C. A. Ayre.
V a r i e t a l d i f f e r e n c e s i n b a r l e y s and m a l t s . Can.
J . R e s e a rc h , C, 16: 456-466.
1938.

5.

Anderson, J . A ., H. R. S a l l a n s ,
and W. 0 . S.
M e r e d i t h . V a r i e t a l d i f f e r e n c e s i n b a r l e y s and
m a l t s . Can. J . R e sea rc h , C, 19: 278-291. 1941.

6.

A s s o c i a t i o n o f O f f i c i a l A g r i c u l t u r a l Chem ists.
Methods of a n a l y s i s . 1935.

7.

B l i s h , M. J . and R. M. S a n d s t e d t . An improved
method f o r t h e e s t i m a t i o n o f f l o u r d i a s t a t i c
v a l u e . C e r e a l C hem istry 10: 189-202. 1933.

8.

B l i s h , M. J . , R. M, S a n d s t e d t , and H. P l a t e n i u s .
C o r r e l a t i o n between d i a s t a t i c power o f f l o u r and
c r u s t c o l o r i n t h e t e s t l o a f and i t s s i g n i f i c a n c e .
C e r e a l C hem istry 6: 121-127. 1929.

9.

Coleman, D. A. R e p o rt o f t h e m a lt a n a l y s i s
s t a n d a r d i z a t i o n committee. C e r e a l Chem, 13:
669-674. 1936.

10.

Eagedorn, E. C., and B. N. J e n s e n . Zur
mikrobestimmung des b l u t z u c k e r s m i t t e l s
f e r r i c y a n i d . Biochemische Z e i t s c h r i f t 135:
4 5 - 4 8 . 1923.

11.

E an e s, C. S. An a p p l i c a t i o n of t h e method o f
Eagedorn and J e n s e n t o t h e d e t e r m i n a t i o n of
l a r g e r q u a n t i t i e s of reducing sugars.
B io c h e m ic a l J o u r n a l 23: 99-106. 1929.

40

12.

H a r r i s , R. H . , and 0, J . B a n a s i k .
E ffects of
e n v iro n m e n t, v a r i e t y , and se aso n on b a r l e y
q u a l i t y . C e r e a l Chem. S9: 148-155. 1953.

13.

Hopkins, R. H . , and C. B. K rause.
B io c h e m is tr y
a p p l i e d t o m a l t i n g and brew ing. D. Van N o s tr a n d
Company, I n c . , New York, 1937.

14.

L e je u n e , A. J . and J*. H. P a r k e r .
Farmers and
s h i p p e r s gu id e t o premiums f o r h ig h q u a l i t y
m a l t i n g b a r l e y . Midwest B a r l e y Improvement
A s s o c i a t i o n , Milwaukee, W isconsin. J a n u a r y , 1953.

15.

L e je u n e , A. J . , W. W. S i s l e r , 0 . J , B anasik, and
R. H. H a r r i s . Methods of b r e e d i n g new m a l t i n g
b a r l e y v a r i e t i e s . The Brewers D i g e s t 26: 57-63.
1951.

16.

M e r e d i t h , W. 0 . S . , H. R, S a l l a n s , and H. Rowland.
P r e d i c t i o n o f m a lt d i a s t a t i c power of h y b r i d
b a r l e y s . S c i e n t i f i c A g r i c u l t u r e 22: 761-771.
1942.

17.

R o b e rtso n , D. W., G. A. Wiebe, and F. R. Immer.
A summary o f l i n k a g e s t u d i e s i n b a r l e y . J o u r ,
Amer. Soc. Agron, 33: 47-64. 1941.

18.

R o b e r t s o n , D. W., G. A* Y/iebe, and R. G. Shands.
A summary o f l i n k a g e s t u d i e s i n b a r l e y . J o u r .
Amer. Soc. Agron. 39: 464-473. 1947.

19.

Rumsey, L. A. The d i a s t a t i c enzymes of wheat
f l o u r and t h e i r r e l a t i o n t o f l o u r s t r e n g t h .
Am. I n s t . Baking, B u l l e t i n 8. 1922.

20.

S a l l a n s , H. R . , and J . A. Anderson. V a r i e t a l
d i f f e r e n c e s i n b a r l e y s and m a l t s . Can. J .
R e s e a r c h , C, 16: 405-416. 1938.

21.

Sandegren, E . , and W. Klang. B a r l e y amylase
and p r o t e i n a s e . J . I n s t . Brewing 56: 313-318.
1950.

22.

S h earo n , W. H ., and H. E. W e i s s l e r . Brewing
I n d u s t r i a l and E n g i n e e r i n g C hem istry 43:
1262-1271. 1951.

41

23.

T e c h n i c a l Committee and E d i t o r i a l Board of t h e
American S o c i e t y of Brewing C hem ists. Methods
o f a n a l y s i s o f t h e American S o c i e t y of Brewing
C h e m ists. Ed. 5, American S o c i e t y o f Brewing
C h e m ists, Chicago. 1949.

24.

Yoger, E. E . , E. E. Schw aiger, H. 0. L e o n h a rd t,
and J . A. M e rte n . The P r a c t i c a l Brewer. Yon
Hoffmann P r e s s , S t , L o u is . 1946.

25.

Weaver, J . C. American b a r l e y p r o d u c t i o n .
B urgess P u b l i s h i n g C o., M in n e a p o li s . 1950.

26.

Weeks, M. Beer and b rew in g i n America.
P a r s o n s & C o., New York. 1949,

A p p le to n ,