2012 GEM Report: Truman State University,
Annual GEM Cooperator meeting, ASTA, Dec. 4, Chicago, IL
Development and evaluation of specialty starch germplasm utilizing GEM biodiversity to
optimize grain quality, composition, and yield.
Duration 2011 – 2015
Mark Campbell, Jon Beck, Marianne Emery, Danica Borje, Akriti Panthi, Jessica Issleib, Elizabeth Vanover,
Jillian Burke and Deborah Boedeker, Kumud Joshi
General Objectives: The GEM program has been used to demonstrate the value of exotic maize
germplasm by developing parent lines and hybrids with altered starch traits that target current
trends in industry and consumer interests. Until now, all of our germplasm developed at Truman has
relied exclusively on materials from GEM which include breeding crosses composed of either 25% or
50% exotic germplasm, of primarily of Latin American origin, with proprietary lines from private
cooperators (1). Using this material, our efforts have focused on developing two distinct hybrid types
with unique starch properties. The first includes high-amylose corn which has a number of niche
applications, one of which includes its use as a source of resistant starch (RS) having prebiotic
properties with well documented health benefits and functional attributes making it well suited as a
dietary supplement in food fortification (2). The second starch type involves developing a
commercially viable source of a slowly digested starch (SDS) from native (unmodified) starch which
has been studied for its potential applications in controlling blood sugar (3). Although a number of
applications exist, our emphasis is towards an SDS starch having a combination of a lowered
glycemic-index (GI) and RS content for a slowed release of sugar into the blood that could be used
therapeutically for diabetic and glycogen disorder patients suffering from nocturnal hypoglycemia.
Already, GEM materials have provided a source of unique alleles altering starch for these
applications. The diversity of materials also provides an ideal opportunity to identify genetic
backgrounds to overcome decreased yield and grain quality associated with mutations altering starch
structure. In addition to several classic starch mutations including waxy (wx) and amylose-extender
(ae) a novel allele at the starch branching enzyme 1 (sbe1) exists in our material we refer to as
(sbe1::gems67) originating from GEMS-0067 are in our lines. Current grain prices and premiums on
specialty grains require these improvements so that production, milling and marketing can be
economically viable.
This past summer several ongoing studies and continued development of germplasm has taken place.
Highlighted are field studies that demonstrate agronomic potential of Amylomaize lines developed
using only GEM parents under varied environmental conditions. Additionally, some projects
involving collaboration with faculty at Truman, A.T. Still Medical College and ISU have been initiated.
Several projects have been ongoing and require further data analysis and description
Hybrid Evaluation
With the assistance of a private cooperator, a hybrid evaluation of experimental crosses was intended to
be grown this past summer. Unfortunately, parent seed for production of the crossing blocks did not
arrive for the Puerto Rico winter growing season (2011/2012) until April 2012. The same material was
sent for crossing during this winter nursery season (2012/2013) and hybrids will be grown in Ames and
another location for the 2013 summer growing season. Materials were selected based on having been
selected for the recessive amylose-extender mutation (ae) and also having been inbred and at least
preliminarily been selected for starch amylose near or above 25%. In each crossing block, testers were
selected based on identification of a line that has performed best as a parent with good general combining
ability and also displayed good grain quality attributes (lack of rot, minimal kernel shrinkage, etc).
Many of these lines have been further inbred and selected within and, therefore, do not represent the most
current inbred materials.
Test crosses currently being made in Puerto Rico for hybrid evaluation in 2013 - SS tester
29 rows
Male
Male
Male
Male
Male
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Set 1
Set 1
Set 1
Set 1
Set 1
Set 1
Set 1
Set 1
Set 1
Set 1
Set 1
Set 1
Set 1
Set 1
Set 1
Set 1
Set 1
Set 1
Set 1
Set 1
Set 1
Set 1
Set 1
Set 1
Set 1
Set 1
Set 1
Set 1
Set 1
Set 1
Row
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
Amylomaize line
SS
SS
SS
SS
SS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
CHIS775:S1911b-120-1-B-B-B////AR16035:S02-615-1-B-B///GEMS-67 --------Tester
CHIS775:S1911b-120-1-B-B-B////AR16035:S02-615-1-B-B///GEMS-67 --------Tester
CHIS775:S1911b-120-1-B-B-B////AR16035:S02-615-1-B-B///GEMS-67 --------Tester
CHIS775:S1911b-120-1-B-B-B////AR16035:S02-615-1-B-B///GEMS-67 --------Tester
CHIS775:S1911b-120-1-B-B-B////AR16035:S02-615-1-B-B///GEMS-67 --------Tester
9353-01/97_DK888N11F2S3_7451-17-b-b/////CH05015:N1204-57-1-B-B////DKXL370B-B-SIB///GEMS-67
BARBGP2:N08a18-332-001-B-B-B/////CH05015:N1204-57-1-B-B////DKXL370:N111-1B-B-SIB///GEMS-67
FS8B(T):N11a-087-001-b-b-sib-b-b/////CH05015:N1204-57-1-B-B////DKXL370:N11a20BB-SIB///GEMS-67
CHO5015:N1204-057-001-b-b-b/////AR03056:N09-24-1-B-B-B////DKXL370:N11a20-31B-SIB///GEMS-67
DKXL370:N11a20-199-002-B-B-B-Sib/////AR03056:N09-182-1-B-B-B////DKXL370:N11-B-SIB///GEMS-67
BR51675:N0620-033-001/////AR03056:N09-24-1-B-B-B////DKXL370:N11a20-31-1-B-B-SIB///GEMS-67
MDI022:N21-B-002-003///// DKXL370:N11a20-234-2-B-B-B////DKXL370:N11a20-31-1-B-SIB///GEMS-67
CH0515:N1502-086-001-b-b-b/////UR13085:N0215-14-1-B-B-B-B////DKXL370:N11a20-31-SIB///GEMS-67
DK888:N11-B-027-001-B-001/////DREP150:N2011d-624-1-B-B////DKXL370:N11a20-1-1B-SIB///GEMS-67
DREP150:N2011d-624-1-B-B////DKXL370:N11a20-31-1-B-B-SIB///GEMS-67
CHO5015:N1204-057-001-b-b-b/////UR13085:N0215-14-1-B///GEMS-67
9353-01/97_DK888N11F2S3_7451-17-b-b////UR13085:N0215-14-1-B///GEMS-67
BARBGP2:N08a18-332-001-b-b-b/////UR13085:N0215-14-1-B///GEMS-67
BR51675:N0620-033-001////UR13085:N0215-14-1-B///GEMS-67
CL-G1607(CML420):N11-008-001-007////UR13085:N0215-14-1-B///GEMS-67
CH0515:N1502-086-001-b-b-b////UR10001:S1813-257-1///GEMS-67
CH05015:N1502-086-001-B-B-B/////AR03056:N09-182-1-B-B-B////CH05015:N15-3-1-B-B///GEMS-67
CH05015:N1502-086-001-B-B-B/////AR03056:N09-24-1-B-B-B////CH05015:N15-3-1-B-B///GEMS-67
DK212T:N11a12-191-001-B-B-B/////AR03056:N09-182-1-B-B-B////CH05015:N15-3-1-B-B///GEMS-67
DK212T:N11a12-191-001-B-B-B/////AR03056:N09-24-1-B-B-B////CH05015:N15-3-1-B-B///GEMS-67
DKXL370:N11a20-199-002-B-B-B-Sib/////AR03056:N09-24-1-B-B-B////CH05015:N15-3-1-B-B///GEMS67
BR51403(PE001):N16-B-044-004-001-001C-001////CH05015:N15-3-1-B-B///GEMS-67
DKXL370:N11a20-199-002-B-B-B-Sib////FS8A(S):S09-362-1-B///GEMS-67
DKXL380:N11-B-007-010-B-002/////CHRIS775:S1911b-120-1-B-B-B////CUBA164:S2012-1-B///GEMS-67
Table 1. Test crosses currently being made in Puerto Rico for hybrid evaluation in 2013 - NS tester
37 rows
Male
Male
Male
Male
Male
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Females
Set 2
Set 2
Set 2
Set 2
Set 2
Set 2
Set 2
Set 2
Set 2
Set 2
Set 2
Set 2
Set 2
Set 2
Set 2
Set 2
Set 2
Set 2
Set 2
Set 2
Set 2
Set 2
Set 2
Set 2
Set 2
Set 2
Set 2
Set 2
Set 2
Set 2
Set 2
Set 2
Set 2
Set 2
Set 2
Set 2
Set 2
Set 2
Row
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
NS
NS
NS
NS
NS
SS
SS
SS
SS
SS
SS
SS
SS
SS
SS
SS
SS
SS
SS
SS
SS
SS
SS
SS
SS
SS
SS
SS
SS
SS
SS
SS
SS
SS
SS
SS
SS
AR03056:N09-191-001-B-B-B-Sib/////AR03056:N09-182-1-B-B-B////CH05015:N15-3-1-B-B///GEMS-67
AR03056:N09-191-001-B-B-B-Sib/////AR03056:N09-182-1-B-B-B////CH05015:N15-3-1-B-B///GEMS-67
AR03056:N09-191-001-B-B-B-Sib/////AR03056:N09-182-1-B-B-B////CH05015:N15-3-1-B-B///GEMS-67
AR03056:N09-191-001-B-B-B-Sib/////AR03056:N09-182-1-B-B-B////CH05015:N15-3-1-B-B///GEMS-67
AR03056:N09-191-001-B-B-B-Sib/////AR03056:N09-182-1-B-B-B////CH05015:N15-3-1-B-B///GEMS-67
2011-01_SE32_S17_F2S4_9148/////CHIS775:S1911b-120-1-B-B-B////AR16035:S02-6B-B///GEMS-67
2011-01_SE32_S17_F2S4_9148-Blk22/00/////CUBA164:S1511b-325-B-B////AR16035:S02-6///GEMS-67
CUBA164:S1511b-325-1-B-B////AR16035:S02-615-1-B-B///GEMS-67
CUBA164:S2012-966-1-B-B////AR16035:S02-615-1-B-B///GEMS-67
DKB844:S1601-073-001-B-B-B-B-B/////CUBA164:S1511b-325B-B////AR16035:S02-615-B-B///GEMS-67
BR105:S1612-008-001-B-B/////DKB844:S1601-73-1-B-B-B////AR16035:S02-615-1-B-B///GEMS-67
DKB844:S1601-73-1-B-B-B////AR16035:S02-615-1-B-B///GEMS-67
2088-01_DK212T_S11_F2S4_9157-Blk29/0ib-/////DKB844:S1601-73-1-B////AR16035:S02-6B///GEMS-67
2011-01_SE32_S17_F2S4_9148-Blk22/00////AR16035:S02-615-1-B-B///GEMS-67
GUAT209:S13 08a-120-001-B-B////CHIS740:S1411a-783-2-B-B////AR16035:S02-615-1-B-B///GEMS-67
2011-01_SE32_S17_F2S4_9148-Blk22/00-sib///GEMS-67
CHIS775:S1911b-120-001-B-B-B-B////2011-01_SE32_S17_F2S4_9148-Blk22/00-sib///GEMS-67
2011-01_SE32_S17_F2S4_9148-Blk22/00////2011-01_SE32_S17_F2S4_9148-Blk22/00-sib///GEMS-67
CUBA164:S2012-444-001-B-B-Sib////2011-01_SE32_S17_F2S4_9148-Blk22/00-sib/// GEMS-67
CHIS740:S11411a-783-002-b-b-b////2011-01_SE32_S17_F2S4_9148-Blk22/00-sib///GEMS-67
CUBA164:S2012-444-1-B///GEMS-67
CHRIS775:S1911b-120-1-B-B-B////CUBA164:S2012-444-1-B///GEMS-67
CUBA164:S1511b-325-001-B-B-B-B-B-Sib/////CHRIS775:S1911b-120-1-//// CUBA164:S20///GEMS-67
FS8A(S):S09-362-1-B///GEMS-67
2011-01_SE32_S17_F2S4_9148-Blk22/00////FS8A(S):S09-362-1-B///GEMS-67
CHIS740:S11411a-783-002-b-b-b/////CUBA164:S1511b-325-1-B-B////FS8A(S):S09-362-1-B///GEMS-67
BVIR155:S2012-029-001-B-B////UR10001:S1813-257-1///GEMS-67
CHIS740:S11411a-783-002-b-b-b////UR10001:S1813-257-1///GEMS-67
DKB844:S1601-073-001-B-B-B-B-B////UR10001:S1813-257-1///GEMS-67
UR10001:S1813-257-1///GEMS-67
2011-01_SE32_S17_F2S4_9148-Blk22/00////UR10001:S1813-257-1///GEMS-67
UR11003:S0302-1011-001-b-b-b////UR10001:S1813-257-1///GEMS-67
BR51675:N0620-033-001////UR10001:S1813-257-1///GEMS-67
BR52060:S0210-143-001-b-b-b////UR10001:S1813-257-1///GEMS-67
GUAT209:S13 08a-120-001-B-B////UR10001:S1813-257-1///GEMS-67
CUBA164:S2008c-289-001-B-B/////AR03056:N09-24-1-B-B-B////DKXL370:N11a20-31///GEMS-67
H99 ae///GEMS-67
Hybrid Evaluations under varying population densities – Ames, IA, 2012
An alternative yield trial/experiment was pulled together and established for the summer of 2012 near Ames,
Iowa. Remnant seed from previous experiments were used to investigate the influence of population density on
yield and agronomic traits of plants and grain for experimental Amylomaize VII hybrids. A study by So et al (2009)
demonstrated that plant architecture can play a role in sweet corn varieties which is likely explained by a
substantial interaction of variety x planting density in this specialty crop. Thirteen hybrids that were made from
GEM x GEM Amylomaize VII were seeded in a split-plot design experiment with population densities (55
seeds/plot vs. 75 seeds/plot) in replicated blocks consisted of the collective plots of hybrids serving as the serving
as the main plots. The density treatment imposed was equivalent to 24,000 plants/per acre (55 seeds/plots ) and
32,670 plants per acre (75 seed/plot). Within each main plot were twelve experimental Amylomaize hybrids
including a commercial Amylomaize VII hybrid check that served as the sub-plots. The experiment suffered from
an exceedingly hot and dry summer. In addition, substantial lodging occurred throughout. Seed emergence was
variable and being several years old (2008) viability was low but the effect of genotype stand% was large. Results
are shown in Table 2 where a crude statistical analysis was performed which requires appropriate correction for
this design. The experiment appeared to display variation from many effects and the results are rather mixed.
This small study did not reveal significant influences on density; however they are not conclusive and future
studies should be pursued since upon examination of the data it suggested that optimal yields with specialty corn
may require closer attention to population density. The highest yield was seen in the hybrid
CHIS775:S1911b////AR16035:S02///GEM67 X DREP150:N2011d////DKXL370:N11a20///GEM67 planted at the higher
density. Overall, the average of the lower densely planted Amylomaize hybrids showed a slightly higher yield and
test weight.
So, Y.F., Williams, M.M. II, Pataky, J.K. 2009. Wild-proso differentially affects canopy architecture and yield components
of 25 sweet Corn Hybrids. HortScience. 44:408-412.
Figure 1. Parent lines in nursery display a wide variety of plant leaf structures and overall plant architectural
differences which may influence their response to planting density.
Table. 2. Yield and agronomic performance of 12 Amylomaize VII hybrids during 2012 in Ames Iowa. Plant seeded at 55 versus75 seeds per plot.
WT
Y/M
Yield (bu/ac)
Hybrid
source
55
75
55 75
AR03056:N09-24////DKXL370:N11a20///GEM67 X CUBA164:S1511b////AR16035:S02///GEM67
4039
12.1
12.5
4.4 87.2 91.1
4.9 82.2 93.6
5.8 90.9 132.2
3.6 66.3 64.6
3.0 63.6 65.0
4.2 87.9 81.3
3.5 78.4 72.7
4.0 100.7 95.7
3.7 116.2 89.9
3.8 107.8 79.2
2.7 78.9 46.6
4.6 96.2 84.6
4.0
AR03056:N09////DKXL370:N11a20///GEM67 X CHRIS775:S1911b////CUBA164:S2012///GEM67
4030
11.4
11.0
AR03056:N09////DKXL370:N11a20///GEM67 X CUBA164:S2012////2011-01_SE32_S17///GEM67
4034
10.9
10.0
AR03056:N09////DKXL370:N11a20///GEM67 X CUBA164:S2012////2011-01_SE32_S17///GEM67
4009
14.4
13.7
CHIS775:S1911b////AR16035:S02///GEM67 X SCR01:N1310///GEM67
4020
16.4
12.9
CUBA164:S1511b////AR16035:S02///GEM67 X DKXL370:N11a20///GEM67
4035
14.9
10.9
proprietary hybrid
check
10.3
6.1
CHIS775:S1911b////AR16035:S02///GEMS67 X AR03056:N09-24////DKXL370:N11a20///GEMS67
4023
12.7
11.3
4.1
3.9
3.8
3.6
3.0
5.5
3.6
4.2
5.1
5.1
4.8
5.4
ave
12.1
11.4
4.3
AR03056:N09////DKXL370:N11a20///GEM67 X CUBA164:S1511b////AR16035:S02///GEM67
4042
11.4
12.6
CHIS775:S1911b////AR16035:S02///GEM67 X DREP150:N2011d////DKXL370:N11a20///GEM67
4022
13.0
18.7
CHIS775:S1911b////AR16035:S02///GEM67 X DK212T:N11a12-/////CH05015:N1204////DKX L370:N11a20///GEM67
4036
8.8
8.6
CHIS775:S1911b////AR16035:S02///GEM67 X CH05015:N1204////CH05015:N15///GEM67
4040
8.8
9.1
55
75
88.0
83.0
SOURCE OF VARIATION
df
P-value
P-value
P-value
BLOCK
1
ns
0.06
ns
POP. D ENSITY
1
ns
ns
ns
GEN
11
0.03
ns
0.07
Table 2. Continued.
Cont’d
%rtldg
%skldg
%stand
stand
MOIST
TWT
source
55
75
55
75
55
75
55
75
55
75
55
75
4039
12.1
12.5
29.4
26.1
90.9
82.6
50.0
59.5
21.4
21.0
54.4
54.1
4042
11.4
12.6
0.0
6.9
58.2
59.7
32.0
43.0
21.8
19.3
54.2
56.9
4022
13.0
18.7
19.7
2.1
59.1
68.7
32.5
49.5
23.9
22.9
53.2
54.1
4036
8.8
8.6
1.7
8.4
57.3
56.9
31.5
41.0
18.7
18.0
54.1
55.1
4040
8.8
9.1
34.5
4.7
70.9
78.5
39.0
56.5
21.3
23.3
55.3
51.7
4030
11.4
11.0
62.5
0.0
43.6
29.9
24.0
21.5
15.9
20.6
55.9
52.2
4034
10.9
10.0
7.9
3.4
69.1
81.2
38.0
58.5
21.9
21.0
54.2
53.9
4009
14.4
13.7
8.2
6.3
77.3
68.1
42.5
49.0
24.4
24.2
51.6
52.6
4020
16.4
12.9
7.9
3.6
69.1
75.0
38.0
54.0
23.1
24.4
53.4
51.8
4035
14.9
10.9
3.6
1.9
74.5
71.5
41.0
51.5
21.2
20.8
53.9
53.1
check
10.3
6.1
7.6
3.4
71.8
60.4
39.5
43.5
16.5
17.3
54.7
43.8
4023
12.7
11.3
2.5
0.0
77.3
61.1
42.5
44.0
17.7
18.4
54.6
52.3
12.1
11.4
15.5
5.5
68.3
66.1
37.5
47.4
20.6
20.9
54.1
52.6
SOURCE OF V ARIATION
df
P-value
P-value
P-value
P-value
P-value
P-value
BLOC
1
ns
0.04
ns
ns
0.00
0.04
POP . DENSITY
1
ns
ns
ns
0.00
ns
ns
GEN
11
0.03
ns
0.00
0.00
0.00
ns
In addition, several experimental hybrids were included in the summer 2012 nursery. The entries were F1 crosses
between several synthetics, Nepal accessions, new GEM releases and a mutant stock possessing sh2-:rev2. There are
materials which have been crossed on to existing Amylomaize VII lines in order to see how the ae allele and modifiers
behave regarding starch properties and agronomic performance. The ears from these F1 plants will segregate normal to
ae kernels in the expected 3:1 ratio so yields will be more representative of the normal endosperm.
The QPM synthetics are of particular interests since recent studies have implicated deficiencies of essential amino acids
in regulating healthy microflora. In a recent article in Nature amino acids,’ The building blocks of protein, into your body
from the food you digest’
PI 586689 incorporates germplasm from CIMMYT
Pool 33 QPM (modified opaque-2) and synthetic
BSCB1(R)C11. Developed by Dr. Brent Zehr at
Purdue University.
(A: F1 ears: PI 586689 x NS amylomaize VII line)
PI 586690. Incorporates germplasm from CIMMYT
Pool 33 QPM (modified opaque-2) and synthetic
BSSS(R)C11. Developed by Dr. Brent Zehr at
Purdue University.
(B: F1 ears: 586690 x SS amylomaize VII line)
Nature: Volume 487 Number 7408 pp405-524 26 July 2012 ACE2 links amino acid malnutrition to microbial ecology and intestinal
inflammation Tatsuo Hashimoto, Thomas Perlot, Ateequr Rehman, Jean Trichereau, Hiroaki Ishiguro+ et al.
Population/GEM release/Synthetic x
GEM Amylomaize VII
WT
Yield
(bu/ac)
MOIST
Y/M
TWT
%
stand
%
skldg
%
rtldg
1
10.7
77.3
21.3
3.6
54.5
76.4
26.2
26.2
1
13.7
98.7
21.7
4.6
55.5
127.3
19.2
21.6
14.0
104.6
18.9
5.6
54.6
100.0
3.3
13.1
Source
Nepal 1701 -- PI 511578 x AR03056:N09B////CH05015:N15///GEMS-67
11:78/493
Nepal 1701 -- PI 511578 /CH05015:N15B///GEMS-67
11:78/498
Nepal No. 1 -- PI 208468 x FS8A(S):S09///GEMS-67
11:70-1/386
1
Nepal 2402 -- PI 511581 x CHIS740:S11411a////2011-01_SE32_S17///GEMS-67
11:80-2/375
1
13.4
101.6
17.4
5.9
57.2
79.1
23.4
24.5
Nepal 5503 -- PI 511601 x 2088-01_DK212T_S11/////DKB844:S1601////AR16035:S02///GEMS-67
11:92-1/355
1
5.2
37.7
22.0
1.8
43.0
92.7
12.5
15.5
MAKKAI - Punjab, India PI 164375 - x DKB844:S1601////AR16035:S02///GEMS-67
11:101-4/353
1
14.6
108.2
19.9
5.6
57.6
88.2
13.2
12.1
Nepal 4109 -- PI 511597 x CHIS740:S1411a////AR16035:S02B///GEMS-67
11:90-1/359
1
8.0
58.2
21.0
2.9
55.0
86.8
4.5
3.4
Nepal 5603 -- PI 511602 x CH05015:N1502/////AR03056:N09////CH05015:N15///GEMS-67
11:93-1/501
1
9.1
65.1
22.6
2.9
55.2
74.3
11.4
20.3
11:90-2/371
1
9.5
68.6
21.7
3.3
54.0
84.0
23.0
5.2
11.3
76.6
26.4
3.0
52.9
81.2
5.3
11.0
Nepal 4109 -- PI 511597 x 2011-01_SE32_S170////2011-01_SE32_S17b///GEMS-67
Nepal 5603 -- PI 511602 x DKB844:S1601////AR16035:S02///GEMS-67
11:93-1/353
1
PI 208472s (POKHARA) -- x DK212T:N11a12B/////AR03056:N09B////CH05015:N15B///GEMS-67
11:74-1/506
1
13.5
99.5
19.8
5.1
55.6
60.4
7.6
1.5
Nepal 3905A -- PI 511591 x DKB844:S1601////AR16035:S02///GEMS-67
11:87-1/354
1
9.3
64.9
25.1
2.7
52.4
72.2
31.6
21.2
1
13.8
98.6
22.3
4.5
56.9
72.9
1.0
5.8
Nepal 4104 -- PI 511595 x DKB844:S1601////UR10001:S1813///GEMS-67
11:89-2/396
2011 GEM Release MBRC10:S1741-B-057 x SCR01:N1310B///GEMS-67
11:67-6/523
2
14.8
118.4
13.4
8.9
54.8
86.4
2.2
2.1
2011 GEM Release (GEMS-0113/GEMS-0091) x CHIS740:S11411a////UR10001:S1813///GEMS-67
11:65/393
2
12.6
96.2
17.5
5.7
57.1
87.3
9.8
27.5
2011 GEM Release MBRC10:S1741-057 x CHIS775:S1911bB////2011-01_SE32_S17///GEMS-67
11:67-6/370
2
14.2
115.2
12.3
9.5
58.3
77.3
2.4
1.3
2
14.4
105.5
20.3
5.2
56.0
61.8
15.1
17.9
2011 GEM Release CUBA164:S2012-444-001-B wx x SCR01:N1310B///GEMS-67
11:66-1/524
PI 586690 HQPSSS - IN, US x BR105:S1612/////DKB844:S1601B////AR16035:S02///GEMS67
11:107-2/348
3
16.0
123.6
16.0
7.6
55.3
91.8
24.6
1.1
PI 586690 - - HQPSSS - IN, US x 2011-01_SE3S17/////DKB844:S1601////AR16035:S02///GEMS67
11:107-1/349
3
15.5
112.4
21.1
5.4
54.8
97.3
16.4
8.3
3
11.2
85.0
17.9
4.8
54.5
73.6
43.2
5.6
PI 586690- HQPSSS -IN, US x AR03056:N09////DKXL370:N11a20B///GEMS-67
11:107-10/443
PI 586689 - HQPSCB - IN, USs x 2011-SE32_S17/////DKB844:S1601////AR16035:S02//GEMS67
11:106-3/349
4
17.0
122.1
21.7
5.6
55.7
84.5
16.3
11.8
PI 586689- HQPSCB -IN, US x BR105:S1612/////DKB844:S1601////AR16035:S02///GEMS-67
11:106-6/351
4
11.7
88.4
18.0
4.9
54.7
59.7
13.1
0.0
PI 586689- HQPSCB -IN, US x BR105:S1612/////DKB844:S1601////AR16035:S02///GEMS-67
11:106-1/351
Florida StaySweet' sh2:rev 2 x DK844:S1601-3-2-B GF0304/GEMS-67
11:111-4/174
4
14.1
103.7
20.0
5.3
55.1
68.7
36.8
8.4
5
8.3
62.7
18.3
3.5
55.4
70.8
34.5
15.5
0.04
0.00
0.00
0.00
0.02
ns
ns
ns
0.01
ns
ns
ns
0.02
ns
Entry
P-value (0.05)
0.05
Block
P-value (0.05)
ns
Development of a bioassay as a possible selection tool for determining prebiotic
efficacy in experimental RS lines and hybrids :
This summer Kumud Joshi spent time working with Dr. S. Hendrich (ISU) and Dr. Vineet Singh,
Professor of Microbiology and Immunology (ATSU) in an effort to develop a rapid screening method to
identify grain samples that likely have high RS values and to subject them to a simulated anaerobic
fermentation environment in order to look at rate of growth of one of the important probiotic
bacterial for which amylomaize VII enhances
Variation in Prebiotic Properties of Experimental Maize Lines with Higher Levels of Resistant Starch
A recessive mutation of a secondary copy of starch branching enzyme, sbe1::gm67, was found at
Truman state from a Guatemalan strain that elevated starch amylose over 70% with resistant starch
increasing to 45% (Campbell et al, 2007). The RS of high amylose starch is a fermentable fiber that
upon entering the lower intestine serves as substrate for probiotic microbial fermentation and
production of short chain fatty acids. Short chain fatty acids such as butyrate, propionate and acetate
provide energy for colonocytes (cells lining the colon). In addition to nutritional effects, butyrate has
been found to reverse neoplastic changes in the lower intestine and induce apoptosis in damaged cells.
Recent studies (Keenan et al, 2112) show that high amylose starch may improve human health
because less of the linear polymer is digested and absorbed in the small intestine, leaving more to
serve as prebiotic fiber for the probiotic microbial residents of the large intestine. Lower amylose
starch has been found to contribute to obesity by influencing long-term energy balance in the body,
and is also associated with colon cancer and diabetes (Le Leu, 2007). Furthermore, clinical trials in
countries with compromised water quality have shown that prebiotic RS maize supplements was
associated with more rapid recovery of patients afflicted with diarrheal diseases. Thus, RS also has
potential to improve Oral Rehydration Treatments (ORT) worldwide. The purpose of the study was to
assess the prebiotic activity of three experimental maize lines by assessing the growth of four strains of
common probiotic bacteria.
The experimental corn lines had 70% amylose content or greater and were developed by traditional
breeding methods at Truman State University. The growth of bacteria on the three experimental lines
was compared to that for a control line with normal amylose content. Four corn lines were predigested in
vitro to mimic human illegal digestion using pancreatin and HCl for 2 h., oven dried for 2 d., and sterilized by
autoclave. MRS growth medium was used to culture bacterial growth. Sterile, predigested corn samples were
added to MRS media with and without glucose. Anaerobic chamber was used to adapt growth medium and
inoculated bacteria. Four anaerobic probiotic bacteria were evaluated for growth on predigested amylomaize
corn using turbidity (Absorbance 610nm) as the growth indicator.
Gram staining was used to assess purity of cultures after growth studies with digested corn
Bacterial Strains (all gram positive rod shape):
• Clostridium butyricum ; Strictly anaerobic, butyric acid producer
• Bifidobacterium longum ; Anaerobic, maintain a normal intestine, boost up immune system
and inhibit growth of gram negative bacteria ??
• Bifidobacterium bifidum ; Anaerobic, aid in vitamin synthesis, improve immunity and
digestion, help in diarrhea treatment, cancer prevention and alcoholic liver restoration
• Lactobacillus sakei ;Facultative anaerobe, hetero-fermentative, commonly found living on
meat and fish
Results: Currently, RS maize is commercially produced using proprietary varieties that limit
widespread production and availability. A long term outcome of this project is to develop RS maize
that can supplement breakfast products such as corn flakes, to help manage risks for colon cancer, high
cholesterol, high blood pressure, and obesity associated with highly refined and processed foods.
Figure 1. B. bifidum growth on MRS (sans
glucose) in presence/absence of predigested
corn starch. Results from one study suggest
that GEM0067 supported more net growth
than the Normal corn.
Figure 2. B. bifidum grew better on the
prebiotic nutrient broth MRS than did C.
butyricum or L. sakei. B. longum did not grow
well on MRS.
Figure 3. Three of the four probiotic bacteria
grew well on Fluid Thioglycollate (FT) broth, a
standard laboratory medium for anaerobic
culture, in this study. In contrast to MRS broth,
B. longum did grow well on FT medium (data not
shown).
A rapid in vitro bio-assay would assist plant breeders screen for high level RS maize for production of
affordable, locally adapted varieties to serve as additives to Oral Rehydration Treatments (ORT) in
countries where diarrheal diseases are common.Selection for increased amylose starch has proven
effective as a primary selection parameter. Other grain components also influence probiotic microbial
populations including the formation of insoluble salts of zinc and magnesium by phytic acid (inositol
hexakisphosphate), which can vary widely among varieties. Additionally, amino acid profiles have
recently been found to play a profound role in gut microflora, particularly tryptophan, which is
notoriously low in maize. Quality Protein Maize (QPM) populations already have been introgressed
into high amylose corn which could improve its efficacy as a prebiotic nutrient to sustain a healthy gut
flora.
•
•
•
•
Campbell, M., Jane, J., Pollak, L.M., Blanco, M.H., O'Brien, A. 2007. Registration of maize germplasm line
GEMS-0067. Journal of Plant Registrations. 1:60-61.
Vineyard, M. L. , and Bear, R. P. 1952 Maize Genetics Coop. News Lett., 26, 5
Keenan MJ, Martin,l RJ, Raggio, AM, McCutcheon KL, Brown IL, Birkett A, Newman SS, Skaf J, Hegsted M,
Tulley R., Blair, E. Zhou J. 2012. High-Amylose Resistant Starch Increases Hormones and Improves
Structure and Function of the Gastrointestinal Tract: A Microarray Study. J Nutrigenet Nutrigenomics
2012;5:26-4
Le Leu, RK. Brown, IL. Hu 2007. Suppression of azoxymethane-induced colon cancer development in rats
by dietary resistant starch. Cancer Biol Therapy 6: 1621–26
Data management work in partnership with Truman Math and Computer science
During this past summer, GEM activities at Truman partnered with Truman States MathBIO program which has
been funded over the past five years with one year remaining. The objective of the NSF funded grant is to
develop collaborations between Math and computer science faculty and students with those in the life sciences.
This project is an insightful effort driven much by Match and Computer science faculty in order to meet industry
demands for people trained in bioinformatics and with knowledge of life science especially in agriculture and
health. This past summer I worked with Dr. Jon Beck where the focus of the project was to create a searchable
data base for unique phenotypic data from the specialty starch breeding activity. The student involved
consisted Deborah Boedeker, Jillian Burke who were involved in initial work in the development of
bioinformatics programs and continued collection of marker data were accomplished.
Creating and Using Phylogenetic Tree Browsers in Marker-Assisted Gene Selection Studies
This project builds on work of Germplasm Enhancement of Maize (GEM) project. The GEM mission is to
demonstrate the value of the naturally occurring biodiversity found in our world’s most traded feed grain [4]
and to incorporate unadapted tropical genetic material into maize lines for commercial use. To this end, Dr.
Campbell and associates have bred a high-amylose maize line, GEMS-0067, that contains genetic material from a
Guatemalan maize line as well common, commercial inbred lines [1]. The aim of this MathBio project is to
compile the data collected by the Campbell laboratory as part of the GEM project into a database and use this
as the basis of an interactive, computerized phylogenetic tree browser interface. Important features of this
system will be to allow for cross-generational comparison of data on several maize lines as well as the
comparison of data of different types, including phenotypic and genotypic data, quickly and efficiently. Principle
functions of the browser will be to confirm the predictions made from the pedigree of the GEMS-0067 maize
line, assist in the creation of further studies of GEMS-0067 through marker-assisted selection, and determine
previously undetected patterns in the characteristics and inheritance of the high-amylose phenotype in GEMS0067.
Figure 1. One example application of the database is the family tree graph shown below that tracks the lineage of certain rows back to
their beginnings. The numbers represented in the nodes of the graph are simply that particular row’s identification number in the
database, whereas the names in the graph are the canonical names of the lines.
Background: Commercial agricultural practices in the United States demand highly inbred crop lines, yet the
practice of inbreeding leads to lessened genetic diversity. This lack of diversity creates a vulnerability to cropfailure episodes, such as those caused by parasitic organisms or droughts. Not only does the lack of genetic
diversity hold the potential for catastrophic problems in US agriculture and economy under certain
circumstances, it also poses a problem for the refinement of beneficial traits and novel applications of plant
products as there is limited genetic material upon which to draw. Past research at Truman State University has
shown an example of the positive effects of diversity by combining traditional breeding techniques with markerassisted selection to establish a new, high-amylose producing maize line, GEMS-0067 [1]. GEMS-0067 is
descendant from H-99 and Oh-43, both commercially-bred US lines, and GUAT209, a native landrace of maize
from Guatemala. Some progeny of the GEMS-0067 line, when crossed with the commercial inbred H-99, gave
markedly greater amylose levels [2]. Amylose is a form of starch that is resistant to digestion in the gut, and
therefore a high-amylose producing maize has potential commercial valuable, with applications in human and
animal health and nutrition. A primary aim of this research is to improve and expand the usage of genotypic
data in future studies involving GEMS-0067, including marker-assisted-selection and QTL mapping as a method
for identifying genes involved producing the desired, high-amylose phenotype and fixing those genes into a
commercially-viable maize line [5].
The database is still in preliminary stages but we propose to expand the database to store the records kept over
the last 15 years by Mark Campbell, as well as the data that has been generated by other groups working on the
GEM Project. Upon completion, the database will contain various data types including near-infrared spectra,
phenotypic data such as kernel weight, and genetic-marker data. Additionally a phylogenetic tree will be
constructed based on the database in order to facilitate a quick, concise understanding of the work done that
has been performed on this project. The tree will serve as a data visualization tool to extend the study and
fixation of the high amylose-producing phenotype in maize.
Results: The principle outcome of this project has been to restructure the data storage procedures used by Dr.
Mark Campbell and staff. This aspect of the project is still ongoing. A database has been created that can store
the pedigree data for previous and future years. Along with the database, a web-based graphical user interface
has been created to allow any user to insert new information into the database in the future. New data will now
be entered into the database instead of being stored in individual computer files or as paper hard copies.
Currently the database holds breeding data through the winter of 2005-06 (minus rows 270-339 of 2003
summer, rows 41-45 of 2004 summer, and rows 444-449 of 2005 summer due to missing data from the Excel
files).
Discussion/Future Research: There are many aspects of the database that could be expanded or improved. An
interface needs to be made to make the data search-able and accessible to Dr. Campbell and his and
collaborators, who intend to use the data to aide further research. The database also needs to be expanded
structurally to include the phenotypic and genotypic data that has been collected along with all data that stems
from future studies. Once this data is all integrated into the database system it will be possible to design more
graphical interfaces and visually represent the data. The use of these new methods of storage and visualization
has great potential for aiding future studies. The process of writing a scientific publication from the data, such as
a germplasm release, will be greatly streamlined with the use of the database, as will the designing of a
quantitative trait loci (QTL) analysis of the GEM-0067 line. In the QTL analysis the highest yielding and lowest
kernels of GEM-0067 will be selected and then screened for markers found in QTLs that are known to affect
kernel weight. The selection of a sample population for this study will be greatly expedited by the ability to view
data such as hundred-kernel weights, starch contents, marker data, etc. in a search-able database. Marker-data
generated from the QTL study will be entered into the database so that statistical analysis of the results can be
performed with the intention of identifying a QTL that strongly correlates with improved kernel quality in the
high-amylose GEM-0067 line.
An additional study to further establish the link between high-amylose phenotype and the homozygous mutant
sbe1-gene modifier, as well as to confirm the accuracy of a new method for starch extraction from maize kernels
is already underway. Starch content of various Amylo-IIV maize lines have already been tested for starch
content. Next these lines will be tested for the presence of a modifier gene that was previously demonstrated to
increase amylose levels to 70% of total starch content. The two extraction methods will then be compared
based on which method more closely correlates with the presence of the modifier. Following the completion of
this study the most accurate starch content data will be entered into the database. This data will be potentially
useful in designing future test crosses as well. Finally, a proposed study that calculates the genetic distance
between some of the GEM lines and common commercial inbred lines will attempt to demonstrate the relative
genetic diversity of the GEM material. This study would be carried out by screening lines for of a number of SSR
markers (between 10-200) from the core set established by the American Seed Trade Association [3]. By
calculating the relative number of polymorphic sequences found at these locations each line and comparing it
the sequences from same locations in commercial inbred lines it may be possible to determine if the GEM lines
are more diverse than traditional material. The results of this genotyping study could also be included in the
database and, when combined with the pedigree and QTL results, form the basis of a graphical user interface.
Such an interface would visually display the lineage of selected markers aiding in visual recognition of patterns
of inheritance. Not only will this feature be helpful in predicting future crosses but it will allow other researchers
and non-science public alike to rapidly familiarize themselves with the project.
Sources:
[1] Campbell M.R, Jane J., Pollakc L., Blancod M.,and O'Briena A., 2007, Registration of Maize Germplasm Line GEMS-0067,
Journal of Plant Registrations, Vol. 1 (1), p. 60-61
[2] Hongxin J., Lio J., Blanco M., Campbell M. and Jane J., 2010, Resistant-Starch Formation in High-Amylose Maize Starch
during Kernel Development, Agric. Food Chem., Vol. 58 (13), p. 8043–8047
[3] Kahler, Alex, et. al, 2010, North American Study on Essential Derivation in Maize: II. Selection and Evaluation of a Panel
of Simple Series Repeat Loci, Crop Science, Vol. 50, p. 486-503
[4] Ludwig, Wolfgang, et. al, 2004, ARB: a software environment for sequence data, Nucl. Acids Res., Vol. 32 (4), p. 13631371
[5]United States Department of Agriculture: Economic Research Service, 2012, "Corn: Trade." Edited by Thomas Capehart.
http://www.ers.usda.gov/topics/crops/corn/trade.aspx#world, May 28, 2012.
Publications/Posters
Joshi K, Campbell M, Cooper C, Singh V. 2012. Variation in Prebiotic Properties of
Experimental Maize Lines with Higher Levels of Resistant Starch. Faculty Mentors: Annual
Interdisciplinary Biomedical Research Symposium, November 8, 2012.
Marianne L. Emery, Victoria F. Halfmann, Mark R. Campbell, Faculty Mentor. 2012.
Assessment of Specialty-Starch Corn Selected Using Maize Biodiversity . 2012 Student
Research Conference:25th Annual Student Research Conference. April 21, 2012
Enato A. Esangbedo and Akriti Panthi,. Mark R. Campbell, Faculty Mentor. 2012. Identifying
and distinguishing between single and double mutant corn using Near Infrared Transmittance
Spectroscopy (Poster.) 2012 Student Research Conference:25th Annual Student Research
Conference. April 21, 2012.
Boedker, D., Burke, J., Campbell, M and Beck, J. 2102. Phylogenetic Tree Browsers in
Marker-Assisted Gene Selection Studies, Departments of Agricultural Science and Computer
Science, Truman State University, Kirksville, MO. Undergraduate Research Conference at the
Interface of Biology and Mathematics November 17-18, 2012 University of Tennessee
Conference Center, Knoxville, TN