KANSAS
KANSAS AGRICULTURAL STATISTICS
Room 200, 632 S.W. Van Buren, Topeka, KS 66603, USA.
E.J. Thiessen, Sherri Hand, and Ron Sitzman.
Jagger still most popular. Jagger was the leading variety of wheat seeded in Kansas for the 2002 crop, according to Kansas Agricultural Statistics (Table 1). Accounting for 42.8 % of the wheat in the state, Jagger increased seven points from a year ago and was the most popular variety in seven of the nine districts. Jagger made the biggest gain in the southwest district. The KSU-maintained variety 2137 ranked second over all, with 15.5 % of the acreage. 2137 ranked first in two districts and second in the other seven. Karl and improved Karl moved up to third position and increased 0.3 points from last year. The OSU-maintained variety 2174 moved up to fourth place with 3.1 percent of the acreage. The fifth most popular variety was TAM 110 with 3.0 % of the acreage in the state. TAM 107 moved down to sixth place with 2.9 %. Ike moved down to seventh place, with 2.6 %. Dominator moved up to eighth place, with 2.0 %. The KSU-maintained variety 2163 remained in the top ten with 1.3 %. Back in the top ten is Vista, with 0.9 %. Acres planted with multiple varieties blended together were not included in the rankings by variety. Blends accounted for 11.4 % of the acres planted statewide and were used more extensively in the north central and central parts of the state. Out of the total state acres planted with blends, 96.5 % had Jagger in the blend and 75.8 % had 2137. All hard white varieties accounted for 1.1 % of the state's acreage. Trego was the leading hard white variety, accounting for 0.8 % of the wheat in the state. The majority of the white wheat was planted in the western third of Kansas.
| Variety | % of acreage | Variety | % of acreage |
|---|---|---|---|
| 1. Jagger | 42.8 | 6. TAM 107 | 2.9 |
| 2. 2137 | 15.5 | 7. Ike | 2.6 |
| 3. Karl/Karl 92 | 3.6 | 8. Dominator | 2.0 |
| 4. 2174 | 3.1 | 9. 2163 | 1.3 |
| 5. TAM 110 | 3.0 | 10. Vista | 0.9 |
| Variety | % of acreage | Variety | % of acreage | Variety | % of acreage |
|---|---|---|---|---|---|
| District 10 (Northwest) | District 40 (North central) | District 70 (Northeast) | |||
| Jagger | 27.2 | Jagger | 20.5 | 2137 | 43.3 |
| 2137 | 15.5 | 2137 | 18.3 | Karl/Karl 92 | 25.6 |
| Vista | 9.4 | Karl/Karl 92 | 14.7 | Jagger | 9.9 |
| TAM 107 | 9.1 | Dominator | 6.4 | Dominator | 3.4 |
| Alliance | 4.1 | 2163 | 3.2 | 2163 | 3.2 |
| District 20 (West central) | District 50 (Central) | District 80 (East central) | |||
| Jagger | 18.8 | Jagger | 41.6 | 2137 | 34.8 |
| 2137 | 18.2 | 2137 | 20.3 | Jagger | 28.1 |
| TAM 110 | 15.2 | Dominator | 5.8 | Karl/Karl 92 | 13.4 |
| TAM 107 | 11.3 | Karl/Karl 92 | 4.0 | 2163 | 4.5 |
| Ike | 6.9 | 2163 | 2.1 | Dominator | 4.1 |
| District 30 (Southwest) | District 60 (South central) | District 90 (Southeast) | |||
| Jagger | 32.8 | Jagger | 63.4 | Jagger | 50.7 |
| 2137 | 12.2 | 2137 | 10.8 | 2137 | 27.7 |
| Ike | 11.7 | 2174 | 6.6 | Karl/Karl 92 | 4.6 |
| TAM 110 | 10.9 | Karl/Karl 92 | 1.4 | 2174 | 3.6 |
| TAM 107 | 6.8 | AgriPro Coronado | 1.4 | 2163 | 1.6 |
Publications.
Monthly Crop. Wheat cultivars, percent of acreage devoted to each
cultivar. Wheat quality, test weight, moisture, and protein content
of current harvest. $10.00
Crop-Weather. Issued each Monday, March 1 through November 30 and monthly, December through February. Provides crop and weather information for previous week. $12.00
County Estimates. County data on wheat acreage seeded and harvested, yield, and production on summer fallow, irrigated, and continuous cropped land. December.
Wheat Quality. County data on protein, test weight, moisture, grade, and dockage. Includes milling and baking tests, by cultivar, from a probability sample of Kansas wheat. September.
Each of the above reports is available on the Internet at the
following address: http://www.nass.usda.gov/ks/
Reports available via E-mail and how to subscribe.
A list of all SSO reports that are available via E-mail can be
found on the Internet at http://www.nass.usda.gov/sub-form.htm,
which provides for automated subscribing. The reports are provided
without charge. To subscribe to one or more of the reports listed,
follow the instructions on the automated form.
KANSAS STATE UNIVERSITY
ENVIRONMENTAL PHYSICS GROUP
Department of Agronomy, Waters Hall, Kansas State University, Manhattan, KS 66506-5501, USA.
M. Stanley Liphadzi and M.B. Kirkham.
Since 1976, all of the biosolids from the city of Manhattan, KS, have been injected into the soil to dispose of them and to use them as soil conditioners and fertilizers. Winter wheat has been grown at the Biosolids Farm during this time and continues to be grown at the farm. Long-term applications of biosolids to soil can result in an accumulation of heavy metals. However, the concentrations of heavy metals in the soil at the Biosolids Farm have not been monitored. We wanted to know if the soil contained abnormal levels of heavy metals. Soil samples were obtained from two sites at the Biosolids Farm. One site had received biosolids for 25 years (since 1976) and the other site had never received biosolids (control site). The samples were analyzed for total concentrations of Cd, Cu, Fe, Mn, Ni, Pb, and Zn. The results are shown in Table 1.
| Time of biosolids application to soil | Cd | Cu | Fe | Mn | Ni | Pb | Zn |
|---|---|---|---|---|---|---|---|
| Years | mg/kg | ||||||
| 25 | 0.82 ± 0.15 | 16.7 ± 2.9 | 8,770 ± 1,400 | 167 ± 61 | 8.93 ± 1.94 | 27.2 ± 3.3 | 31.2 ± 2.5 |
| 0 | 0.88 ± 0.27 | 8.5 ± 3.1 | 12,000 ± 3,870 | 212 ± 74 | 12.4 ± 4.5 | 32.6 ± 6.9 | 20.7 ± 7.0 |
| Mean | 0.5 | 20 | Not known | 850 | 40 | 10 | 50 |
| Normal range | 0.01 - 0.7 | 2 - 100 | 200 - 100,000 | 100 - 4,000 | 5 - 5,000 | 2 - 200 | 10 - 300 |
The results show that, after 25 years of application of biosolids to the farm, concentrations of heavy metals have not increased in the soil. The winter wheat at the Biosolids Farm is not being grown on soil contaminated with heavy metals.
We thank Dr. Abdu Durar, Assistant Director of Utilities, Wastewater, City of Manhattan, Kansas, for supplying the soil samples from the Biosolids Farm.
Dr. Fernando Madrid, former postdoctoral student, has returned to Spain to accept the job as director of the analytical laboratory of the Spanish government's Institute of Natural Resources and Agrobiology in Seville.
THE WHEAT GENETICS RESOURCE CENTERDepartment of Plant Pathology, Throckmorton Hall, Kansas State University, Manhattan, KS 66506-5502, USA.
http://www.ksu.edu/wgrc/
B. Friebe, R.G. Kynast, P. Zhang, L. Qi, M. Dhar, and B.S. Gill.
Alien gametocidal chromosomes cause extensive chromosome breakage prior to the S-phase in the first mitotic division of gametophytes lacking the alien chromosome. The broken chromosomes may be healed either by the addition of telomeric repeats in the gametophyte or undergo fusions to form dicentric or translocation chromosomes. We showed that dicentric chromosomes undergo breakage-fusion-bridge (BFB) cycles in the first few mitotic division of the sporophyte, are partially healed before the germ line differentiation regimen, and are healed completely in the ensuing gametophytic stage. The gametocidal factor on chromosome 4Mg of Ae. geniculata was used to induce dicentrics involving the satellite chromosomes 1B and 6B of wheat, T. aestivum. The dicentrics T1BS·1BL-2AL·2AS and T6BS·6BL-T4BL·4BS initiated BFB cycles that ceased 2 to 4 weeks after seed germination. At the end of the BFB cycles, we observed deficient 1B and 6B chromosomes with breakpoints in proximal regions of the 1BL and 6BL arms. The process of chromosome healing was analyzed in root-tip meristems, at meiotic metaphase I, and in the derived progenies by fluorescence in situ hybridization analysis using a telomeric probe pAtT4. The results show that chromosome healing in wheat occurs during very early mitotic divisions in the sporophyte by de novo addition of telomeric repeats and is a gradual process. Broken chromosome ends have to pass through several cell divisions in the sporophyte to acquire the full telomeric repeat length.
S.A. Jackson, P. Zhang, W.P. Chen, R.L. Phillips, B. Friebe, S. Muthukrishnan, and B.S. Gill.
Transformation of plant genomes by biolistic methods has become
routine over the past decade. However, relatively little is known
about how transgenes are physically integrated into the host genome.
Using a high-resolution physical mapping technique, fluorescence
in situ hybridization on extended DNA fibers (fibre-FISH), 13
independent transgenic wheat lines were analyzed to determine
the structural arrangement of stably inherited transgenes in host-plant
chromosomes. Twelve transgenic lines were transformed with a single
plasmid and one line was cotransformed with two separate plasmids,
which cosegregated genetically. Three basic integration patterns
were observed from the fiber-FISH experiments: Type I, large,
tandemly repeated integration; Type II, large tandem integrations
interspersed with unknown DNA; and Type III, small insertions,
possibly interspersed with unknown DNA. Metaphase FISH showed
that the integration of transgenes was in both hetero- and euchromatic,
as well as proximal, interstitial, and distal regions of the chromosomes.
In the transgenic plants, the type of promotor used, rather than
the chromosomal site of the transgene integration, was most critical
for transgene expression. The integration of the transgenes was
not associated with detectable chromosomal rearrangements.
W.L. Li and B.S. Gill.
The Sh2/A1 orthologous region of maize, rice,
and sorghum contains five genes in the order Sh2, X1,
X2, and two A1 homologues in tandem duplication.
The Sh2 and A1 homologues are separated by ~20 kb
in rice and sorghum, and by ~140 kb in maize. We analyzed the
fate of the Sh2/A1 region in large-genome species
of the Triticeae (wheat, barley, and rye). In the Triticeae, synteny
in the Sh2/A1 region was interrupted with a break between X1
and X2 genes. A1 and X2 genes remained collinear
in homoeologous chromosomes as in other grasses. The Sh2
and X1 orthologs
remained collinear but were translocated to a nonhomoeologous
chromosome. X1 gene was duplicated on two nonhomoeologous
chromosomes, and surprisingly a paralog shared much higher homology
than the orthologous copy to the X1 gene of other grasses.
No tandem duplication of A1 homologues was detected but
duplication of A1 on a nonhomoeologous barley chromosome
6H was observed. Intergenic distances expanded greatly in wheat
as compared to rice. Wheat and barley diverged from each other
12 million years ago and both showed similar changes in the Sh2/A1
region suggesting that the break in colinearity as well as X1
duplications and genome expansion occurred in a common ancestor
of the Triticeae species.
E.D. Badaeva, A.V. Amosoma, O.V. Muravenko, T.E. Samatadze, N.N. Chikida, A.V. Zelenin, B. Friebe, and B.S. Gill.
Six polyploid Aegilops species containing the D genome were studied by C-banding and FISH using clones pTa71 (18S-5.8S-26S rDNA), pTa794 (5S rDNA), and pAs1 (noncoding repetitive DNA sequence) as probes. The C-banding and pAs1-FISH patterns of Ae. cylindrica chromosomes were identical to those of the parental species. However, inactivation of the NOR on chromosome 5D with a simultaneous decrease in the size of the pTa71-FISH site was observed. The N^v^ and D^v^ genomes of Ae. ventricosa were somewhat modified as compared with the N genome of Ae. uniaristata and the D genome of Ae. tauschii. Modifications included minor changes in the C-banding and pAs1-FISH patterns, complete deletion of the NOR on chromosome 5Dv, and the loss of several minor 18S-5.8S-26S rDNA loci on N^v^-genome chromosomes. According to C-banding and FISH analyses, the D^cr1^ genome of Ae. crassa is more similar to the D^v^ genome of Ae. ventricosa than to the D genome of Ae. tauschii. Mapping of the 18S-5.8S-26S rDNA and 5S rDNA loci by multicolor FISH suggests that the second (X^cr^) genome of tetraploid Ae. crassa is a derivative of the S genome (section Emarginata of the Sitopsis group). Both genomes of Ae. crassa were significantly modified as the result of chromosomal rearrangements and redistribution of highly repetitive DNA sequences. Hexaploid Ae. crassa and Ae. vavilovii arose from the hybridization of chromosomal type N of tetraploid Ae. crassa with Ae. tauschii and Ae. searsii, respectively. Chromosomal type T1 of tetraploid Ae. crassa and Ae. umbellulata was the ancestral form of Ae. juvenalis. The high level of genome modification in Ae. juvenalis indicates that it is the oldest hexaploid species in this group. The occurrence of hexaploid Ae. crassa was accompanied by a species-specific translocation between chromosomes 4D^cr1^ and 7X^cr^. No chromosome changes relative to the parental species were detected in Ae. vavilovii, however, its intraspecific diversity was accompanied by a translocation between chromosomes 3X^cr^ and 3D^cr1^.
Moha Ferrahi completed his Ph.D. degree in December, 2001, and has returned to Morocco. Li Huang completed her Ph.D. degree in May, 2002. She will remain at the WGRC as a postdoctoral research associate. Vasu Kuraparthy from India is a new Ph.D. student beginning in the autumn of 2001.
USDA-ARS Plant Science and Entomology Research Unit
Throckmorton Hall, Manhattan, KS 66506-5502, USA.
Gina Brown-Guedira, Safarali Naimov (Research Institute of Plant Physiology and Genetics, Academy of Sciences of the Republic of Tajikistan, Dushanbe, Tajikistan), and W.J. Raupp (Wheat Genetics Resource Center, Kansas State University, Manhattan, KS).
The territory of Tajikistan is indigenous to a wide variety of cultivated and wild cereal species. Although the territory was identified as a secondary center of diversity of the tribe Triticeae by N.I. Vavilov, collections of wheat germ plasm from Tajikistan are limited. Collections in the GRIN database of the USDA National Plant Germplasm System consist only of a small number of accessions of T. aestivum (12) and two collections of T. spelta, all of which were obtained from the Vavilov Institute. There are no collections of Aegilops species from Tajikistan in the USDA collection.
An ongoing collaboration between the Academy of Sciences of Tajikistan and the USDA-ARS Manhattan, KS, has led to the recent acquisition of 40 landraces of T. aestivum and 82 collections of wild Aegilops species, including 43 Ae. tauschii, 16 Ae. cylindrica, 16 Ae. triuncialis, and 7 Ae. crassa. Collections were made during August of 2001 by Dr. Safarali Naimov in the central part of Tajkistan, as well as the Hissar and Khanaka mountainous regions and parts of Kyrgystan and Uzbekistan (Table 1). A portion of the collected seed was planted in the field at Dushanbe, Tajikistan, for seed increase and to evaluate the collection for tolerance of high salt concentration in the soil. Preliminary analyses identified a landrace collection of T. aestivum from central Tajikistan tolerant of soil salinity.
A sample of collected seed was sent to the Wheat Genetics Resource
Center at Kansas State University. Collections were assigned accession
(TA) numbers, and seed was planted for increase in the greenhouse
at Manhattan, KS. Seedlings were evaluated for reaction to PNMQ
(avirulence/virulence formula 2a,11,16,17,26/1,2c,3a,3ka,9,10,18,24,30)
and PRTUS6 (2a,3ka,9,61,18,24,26,30/1,2c,3a,10,11,17) isolates
of P. triticina (Table
1). Leaf rust is a serious disease of wheat in many parts
of the world, including most wheat-growing areas of the U.S. and
central Asia. Six wheat landrace collections were resistant or
heterogeneous to both isolates of P. triticina tested.
Leaf rust reactions of Ae. cylindrica and Ae. triuncialis
collections varied with the two isolates. With the exception of
TA10362, all collections of both species had resistant or heterogeneous
reactions when tested with PRTUS 6. Six accessions of Ae. cylindrica
were resistant or heterogeneous when inoculated with the PNMQ
race of leaf rust. All of the Ae. triuncialis collections,
except TA10362, had resistant or heterogeneous reactions to PNMQ.
All collections of Ae. tauschii and Ae. crassa were
susceptible to the leaf rust isolates tested. Species designations
of the collection are being verified and seeds will be sent to
the USDA National Small Grains Collection. Small samples of seed
may be requested from the Wheat Genetics Resource Center. This
work is supported by the U.S. Civilian Research and Development
Foundation for the Independent State of the Former Soviet Union.
U.S. GRAIN MARKETING AND PRODUCTION RESEARCH CENTER
USDA, Agricultural Research Service, Manhattan, KS 66502, USA.
O.K. Chung, F.E. Dowell, S.R. Bean, G.L. Lookhart, J.B. Ohm, M. Tilley, L.M. Seitz, M.S. Ram, D.B. Bechtel, M.E. Casada, S.H. Park, J.D. Hubbard, B.W. Seabourn, M.S. Caley, J.D. Wilson, R.E. Dempster, J.M. Downing, J.E. Throne, J.E. Baker, and C.S. Chang.
D.B. Bechtel and J.D. Wilson.
Plastids in the coenocytic endosperm of young wheat caryopses were mostly in the form of pleomorphic proplastids with a few of the plastids containing small starch granules. Following cellularization of the coenocytic cytoplasm, the endosperm outer layer or two became meristematic and continued to divide until about 14 days-after-flowering (DAF). During the first week of endosperm development, newly divided cells had plastids that were pleomorphic in shape, while subaleurone cells interior to the meristematic region contained amyloplasts that contained a single size class of starch granules (type A starch granules). The pleomorphic plastids exhibited protrusions that extended a considerable distance through the cytoplasm. Amyloplasts in the interior to the meristematic region did not exhibit the protrusions. Both subaleurone and central endosperm cells had amyloplasts that exhibited protrusions at 10-12 DAF, with some protrusions containing small starch granules. By 14 DAF, endosperm amyloplasts lacked protrusions and two sizes of starch, large type A and small type B granules, were present in the cells. Amyloplast protrusions were numerous again at 17 DAF in both subaleurone and central endosperm cells and by 21 DAF a third size class of small, type-C starch granules was present in the cytoplasm. Amyloplasts in the endosperm of wheat apparently divided and increased in number via protrusions, since binary fission typical of plastid division was never observed. Protrusions were observed in the coenocytic cytoplasm, in dividing cells, in subaleurone and central endosperm cells at 10-12 DAF, and in subaleurone and central endosperm cells at 17 DAF. The results suggest that there are three sizes of starch granules produced at specific times during wheat endosperm development.
D.B. Bechtel and J.D. Wilson.
Starch is most abundant storage reserve in the wheat caryopsis
yet little is known about its influence on end use properties.
Starch was isolated from wheat grains of different classes and
analyzed using digital image analysis coupled to a light microscope
to determine starch size distributions. The image analysis data
was converted into volume data. Starch granules with diameters
greater than 5 µm were treated as oblate spheroids for calculating
volume using the formula for an oblate spheroid. The measured
equivalent diameter and an estimated starch granule thickness
value were used for the major and minor axes in the oblate spheroid
formula, respectively. Granules less than 5 µm in diameter
were treated as spheres. Starch granules that had their perimeter
touching the edge of field of view had their volumes corrected
using correction formulae. Correction formulae were developed
for each wheat class or starch size distribution class. Correction
formulae were important because without them up to 50 % of the
large type-A granules could be under counted. Data indicated that
there can be a wide variation in the size distribution of starch
depending on wheat class and environmental effects. Some wheats
exhibited a trimodal distribution of starch while others only
exhibited a bimodal distribution. This data will be used to help
predict wheat quality.
S.R. Bean and G.L. Lookhart.
HPCE is capable of producing high-resolution, rapid separations
of cereal proteins. Furthermore, HPCE has been shown to be highly
reproducible in terms of migration time. However, little work
has focused on the quantitative reproducibility of cereal protein
separations. Several factors such as sample matrix, sample evaporation,
voltage ramp-up time, sample injection time, and capillary end
cut were evaluated for their involvement in quantitative reproducibility.
These experiments showed that preventing sample evaporation, using
optimum injection times, and insuring a clean, square cut on the
capillary all improved the reproducibility of peak areas. Combining
these factors together into an optimized procedure produced reproducibility
with peak areas varying by < 1.76 % RSD. Migration time also
was excellent under these conditions, varying only 0.45 % RSD.
Other variables such as peak area %, peak height, and peak height
% also showed good reproducibility with an RSD < 4%. Increasing
the voltage ramp up time from 0.17 to 0.68 was found to increase
peak efficiency by ~150 %. This factor had no effect on quantitative
reproducibility, however. The gradual buildup of contaminants
on the capillary walls was found to occur overtime and decreased
both separation efficiency and reproducibility. Rinsing capillaries
periodically with appropriate solvents delayed this effect. Peak
efficiency was found to be a good marker for capillary performance
and lifetime.
S.R. Bean and G.L. Lookhart.
The use of multiangle-laser light scattering (MALLS) in conjunction with size-exclusion chromatography (SEC) was investigated for characterizing wheat proteins. Four solvent systems including 50 % acetonitrile/0.1 % trifluoroacetic acid, 50 mM Na2PO4 pH 2.5, 500 mM acetic acid, and 50 mM Na2PO4 pH 7.0 + 1 % SDS were evaluated for protein extraction as well as for use as SEC mobile phases for MALLS analysis. The dn/dc values for wheat proteins were measured in each solvent. All mobile phases except for the SDS solvent showed dn/dc values between 0.16 and 0.20, which were similar to values reported for other proteins. The SDS solvent showed dn/dc values of 0.32, which was similar to that found for other proteins analyzed in the presence of SDS. Although all solvents showed similar resolution when used as mobile phases in SEC analysis, the SDS solvent extracted the most protein (approximately 82 %) in the unreduced form. This solvent system also displayed no concentration dependent or electrostatic effects during MALLS analysis. The SDS soluble and insoluble proteins were characterized by MALLS and Mw distributions ranging up to 1.4 x 107 Da were found for the insoluble proteins. The effect of the column-void volume also was examined as was the effect of sonication on the Mw distribution.
C. Detvisitsakun, W. Zhang, S. Muthukrishnan, G.L. Lookhart, and G.H. Liang.
To improve drought tolerance, a gene encoding a late embryogenesis
abundant (LEA) protein, HVA1, from barley was introduced into
a HRWW Jagger and a HWWW Lakin using biolistic bombardment. The
gene construction, containing HVA1 gene driven by the rice Act1
promoter and the selectable marker gene, bar, under the control
of CaMV35S promoter, was delivered to embryogenic calli. One transgenic
Jagger wheat plant was obtained. This plant survived in medium
containing 5 mg/l ammonium glufosinate during the tissue culture
processes and has normal morphology. The plant tested positive
for the PCR analysis of bar gene and was resistant to 0.1 % (v/v)
herbicide LibertyTM. Southern hybridization analysis showed the
integration of the bar and HVA1 genes into the genome
of this plant. The 27 kD of HVA1 protein also was detected in
this plant as shown by Western hybridization.
M. Tilley, A. Dotson, M. Phillips, G. Liang, and G. Lookhart.
Aegilops tauschii is a valuable genetic resource for improvement of hexaploid wheat. Crosses of the cultivar Century with the Ae. tauschii accessions TA2450 and TA2460 exhibited shorter mixing times and improved milling and baking characteristics when compared to the parental hexaploid line. Novel HMW-glutenin subunits Dx43 and Dy44 were identified in these crosses and have been characterized at the protein and DNA level. A cDNA library was constructed in order to obtain HMW-glutenin subunits cDNA clones for plant transformation. Developing kernels of Ae. tauschii accessions TA2460 and TA2450 were removed from heads at 10-25 days post-anthesis (when glutenin proteins are synthesized and maximal levels of specific mRNA are present). The kernels were frozen in liquid nitrogen and frozen kernels were shipped to Life Technologies (Rockville, MD) for the construction of a cDNA library. The library was titered, plated, and screened using a nonradioactive method that employs a digoxygenin labeled probe. The probe was the 1.8-kb, internal-repeat fragment of the HMW-glutenin subunit Dx43 cloned from genomic DNA using PCR. Approximately 2,000 colonies were screened, and 89 demonstrated significant hybridization to the probe. Plasmid DNA was purified from tentative positives and analyzed by restriction analysis to determine insert size and restriction pattern compared to PCR amplified HMW-glutenin subunits. Several clones were identified and sequencing of cDNA ends verified that cDNAs contained the full coding region of HMW-glutenin subunits Dx 43 and Dy 44.
K.A. Tilley, K.E. Bagorogoza, H. Kwen, and M. Tilley.
Formation of the three-dimensional protein network known as gluten during dough-mixing and bread-making processes is extremely complex. A specific subset of the proteins comprising the gluten complex, the glutenin subunits, directly affects bread-making quality. Glutenin subunits have not been shown to exhibit any definitive structural differences that can be directly correlated to their ability to aggregate into the gluten complex and affect bread-making quality. Evidence presented here indicates that tyrosine bonded species form in wheat doughs during the processes of mixing and baking and are major contributors to the structure of the gluten network. Various oxidizing and reducing agents that have been used in the baking industry directly affect tyrosine bonds. Tyrosine bonds between synthetic glutenin peptides form in vitro under baking conditions in the presence of potassium bromate and in the presence of water-soluble extract of flour. Bond structures and formation during the bread-making processes have been documented by HPLC, NMR, and mass spectroscopic analyses. Flours and doughs from other non-wheat grains have been examined for their abilities to form tyrosine crosslinks. Comparisons of tyrosine crosslinks in soft, hard, and durum wheats have been made and show dramatic differences. The formation of tyrosine crosslinks in developing wheat kernels has also been documented, shedding light on the biological mechanisms for tyrosine crosslink formation. The relevance of these data to wheat quality will be discussed.
M. Tilley, S.R. Bean, P.A. Seib, R.G. Sears, and G.L. Lookhart.
M. Tilley.
The use of GM materials in foods is increasing. Labeling of
such products will be dependent upon methods capable of sensitive
and accurate detection. Detection of transgenic events can be
based upon the detection of the novel proteins or their specific
activities, DNA encoding the specific proteins, or DNA flanking
the coding region sequences (e.g., promoter, terminator). DNA-based
analysis is the method of choice due to the fact that DNA is highly
stable and PCR-based analyses provide very high sensitivity and
specificity. Processing steps have a profound effect upon the
properties of proteins and DNA present in the final product. This
project was designed to examine the effects of the bread-making
process on wheat DNA extracted from various steps in the process.
Samples were taken from wheat kernels, milling fractions, flour,
fully mixed dough, 1st punch, 2nd punch, moulding, pan stages,
during bake (5, 10, and 15 min), and after bake (1, 3, and 5 days).
Total DNA was purified, quantified spectrophotometrically, and
integrity was evaluated on ethidium bromide stained agarose gels.
DNA purified from kernels demonstrated intact high molecular weight
DNA (> 12 kb), whereas that from flour exhibited a broad smear
of DNA ranging from > 12 kb to < 0.3 kb. DNA purified from
bread exhibited a smear with a maximal size of 0.4 kb with an
average size of 0.2-0.3 kb. Samples were utilized in PCR reactions
to amplify products representative of gene sequences present at
different copy numbers within the wheat genome.
Wheat is the major crop representing about one-third of the world grain production. The U.S. produces about 66.5 MMT (2.24 billion bushels) of which 43 % (28.8 MMT) enters the export market. The majority of the wheat is milled into flour for food uses, the remainder is used for animal feed, seed, and industrial uses. Nearly one-half of the wheat produced in the U.S. is HRWW, which is grown in the Great Plains Area. Turkey, was first HRWW grown in the Great Plains in 1873 and today's wheats differ in many ways. The number of HRWW varieties has increased from five in 1919 to 164 in 1984. Important traits selected in HRWW breeding are yield, test weight, kernel characteristics, disease resistance, stress tolerance, and agronomic appearance. Major end-use quality attributes of HRWW are milling and bread-making characteristics. Nearly 100 % of all released HRWW cultivars have been evaluated by the USDA-ARS Hard Winter Wheat Quality Laboratory (HWWQL). HRWW quality shows increasing trends in kernel weight and milling yield, but decreasing trends in protein content. The wheat industry is changing with the introduction of HWWW and additional products. HWWW can yield 12 % more flour with color properties that are desirable for Asian products. The HWWQL has contributed to the HWWW-breeding program by testing the milling and bread-making quality of HWWW varieties. Wheat use in the U.S. is changing from solely white bread to variety breads and nonbread products such as tortillas, pizza crust, and noodles. Future opportunities include wheats developed for niche markets and the role biotechnology will have in wheat improvement. As with white wheat, identification and segregation issues will need to be addressed.
O.K. Chung, J.B. Ohm, G.L. Lookhart, and R.F. Bruns.
Twelve cultivars each of HWW and HSW were grown three crop
years in a unique growing environment in California that allows
for synchronous grain fill of all genotypes thus removing a normally
strong environmental component and allowing a better investigation
of the genetic component differences. Through the 3 years, the
HSW showed significantly higher mean values of protein and gluten
contents, kernel hardness, and loaf volume but lower gluten index
than HWW. Specifically, wheat near-infrared reflectance hardness
score (NIR-HS) overlapped very little among individual cultivars
of the two classes. Therefore, differences in wheat hardness between
HWW and HSW might be caused by genetic background. The HWW and
HSW, grown side by side, could be clearly classified by canonical
analysis, using wheat characteristics including single kernel
parameters in addition to NIR-HS. Principal component regression
analysis indicated that flour yields and loaf volumes could be
estimated using wheat characteristics and/or single-kernel parameters,
showing a good potential for screening early generation breeding
lines.
O.K. Chung, J.B. Ohm, M.S. Caley, and B.W. Seabourn.
Single kernel and mixograph parameters of hard winter wheats
were gathered from federal regional nurseries from 1990 to 1999.
Eight characteristics and 12 machine parameters obtained from
Single Kernel Characterization System were used to develop a prediction
model of flour yield by continuum regression. Flour yield showed
mean values of 68.8 % and standard deviations of 6.6 and 3.5 for
calibration (n = 1,200) and validation sets (n = 300), respectively.
Prediction model of flour yield showed an R2 of 0.696 for calibration
set and 0.684 for validation set. Wheat protein content, single
kernel characteristics, and objective computer-analyzed mixograph
parameters also were used to develop prediction models of bread-making
properties by continuum regression. Bread loaf volume showed mean
values of 880 cm3 and standard deviations of 96 and 86 for calibration
and validation sets, respectively. Prediction model of bread loaf
volume showed R2 of 0.754 and 0.698 for calibration (n = 1,097)
and validation (n = 319) sets, respectively. Prediction models
of mixograph mixing tolerance and baking water absorption, mixing
time, and specific loaf volume showed R2 values of 0.713, 0.726,
0.865, and 0.814 for the calibration set and 0.676, 0.741, 0.835,
and 0.779 for the validation set, respectively.
S.H. Park, O.K. Chung, and P.A. Seib.
Flours with three different protein contents (PC) (9.8 %, 11.9 %, and 13.2 % on a 14 % mb) were used to study the effects of flour particle size (PS) on the experimental bread-making properties including loaf volume (LV) and internal characteristics such as crumb grain, fineness, and elongation ratio. Flour PS was manipulated by two ways: first, three flours were fractionated into three different fractions (< 53 mm, 53-75 mm, and > 75 mm) using sieving machine; and second approach was that three flours were further ground into four different levels of PS (7,000 rpm x 1, 14,000 rpm x 1, 14,000 rpm x 2, 14,000 rpm x 3) using Alpine Pin mill. Medium PS fraction flours (53-75 mm) showed the highest LV and small PS fraction flours (< 53 mm) showed the lowest LV probably because PC of each fraction played a major role. Crumb grain scores determined by baking expert and elongation ratio determined by CrumbScan V 3.0 (American Institute of Baking, Manhattan, KS) were higher for smaller PS fractions except low PC flour. Further ground flours, which could avoid the effect of different PC, showed increases in LV compared to original flours. Flours ground by 7000 rpm x 1 and 14000 rpm x 1 showed the highest elongation ratios and LV, respectively, for 11.9 % and 13.2 % protein flours. Flour PS distribution affected both LV and internal characteristics of experimentally baked pup-loaf breads.
F.M. Dupont, S.B. Altenbach, O.K. Chung, R. Chan, and R. Lopez.
The HRSW Butte 86 and the HRWWs Cheyenne and Arapahoe were
grown in pots under controlled environmental conditions. All three
cultivars have the same complement of HMW-glutenin subunits, including
Dx5 and Dy 10, which contribute to good gluten quality. To understand
the effects of environment on bread-making quality, wheat was
grown under different regimens of fertilizer, water, and daytime
and nighttime temperatures. Post-anthesis fertilizer increased
wheat protein content per wheat grain whereas head and drought
reduced the duration of starch deposition, and thus reduced kernel
weight. In the absence of post-anthesis fertilizer and heat and
drought increased flour protein content. Loaf volume and SDS-sedimentation
volume were highly correlated with flour protein content, regardless
of the environmental treatment. Some mixograph parameters also
were correlated with protein content, regardless of environmental
treatment. The results indicate that flour protein quality for
these wheat cultivars were remarkably stable over a wide range
of protein contents, whether achieved by varying fertilizer, temperature,
or water during grain fill.
O.K. Chung, J.B. Ohm, A.M. Guo, C.W. Deyoe, G.L. Lookhart, and J.G. Ponte, Jr.
Free lipids (FL) were extracted from straight grade flours
(SF) and their air-classified high-protein fractions (ACHPF) of
nine hard winter wheats. The mean values of FL contents in 10-g
(db) SF and ACHPF were, respectively, 92.8 and 178.5 mg for total
FL, 74.1 and 141.9 mg for nonpolar lipids (NL), 12.8 and 20.9
mg for glycolipids (GL), and 4.9 and 12.0 mg for phospholipids
(PL). FL compositions of SF and ACHPF showed nonsignificant difference
in NL (80.7 and 81.1 % of the FL) but significant differences
in GL (13.9 and 12.0% of the FL) and PL (5.4 and 6.9 % of the
FL). Fortification of SF with ACHPF by blending to reach protein
content to 13 % increased protein and gluten quantity and, thereby,
loaf volume but decreased gluten index, loaf volume regression,
and crumb grain scores. NL contents showed significant relationships
with dry gluten contents (r = 0.79) and gluten index (r = -0.83)
values, indicating that high NL content in ACHPF could decrease
gluten quality of fortified flours. Thus, an optimum balance should
be kept when fortification
process is practiced.
J.B. Ohm and O.K. Chung.
Hard winter wheat flours (n = 72) were analyzed for free lipids (FL) and their relationships with quality parameters. The two main glycolipid (GL) classes showed contrary simple linear correlations (r) with quality parameters. Specifically, kernel hardness parameters, flour yields, and water absorptions had significant negative correlations with monogalactosyldiglycerides (MGDG) but positive correlations with digalactosyldiglycerides (DGDG). MGDG showed negative correlations with gluten content but positive correlations with gluten index. The percentages of DGDG in FL had significant positive correlations among cultivars (n = 12) with mixograph and bake mix times (r = 0.71, P < 0.01 and r = 0.67, P < 0.05, respectively), mixing tolerance (r = 0.67, P < 0.05), and bread-crumb grain score (r = 0.71, P < 0.01). These results suggest that increasing DGDG in FL could contribute to enhancing wheat quality attributes including milling, dough mixing and bread-making quality characteristics. FL content and composition (the ratio of MGDG or DGDG to GL) supplement flour protein content to develop prediction equations of mixograph mix time (R2 = 0.89), bake mix time (R2 = 0.76), and loaf volume (R2 = 0.72).
J.D. Hubbard, J.M. Downing, and O.K. Chung.
In recent years interest has shown in developing analytical methods for the extraction of lipids from various cereal grains using a Supercritical Fluid Extraction (SFE) system because of environmental, toxic exposure, and cost effects. The SFE methods are cost effective, less time consuming, and friendlier to the environment. Most recent interest is to apply SFE to extract total lipids from RTE Breakfast Cereals, instead of acid hydrolysis, for nutritional labeling purposes. We searched for a modifying solvent which would, in a binary supercritical mixture, successfully extract the total lipids including starch lipids. We used mixture of 1-propanol-water (3:1) 40 % by volume with carbon dioxide to form a binary supercritical fluid at 10,000 psi, 120°C, and 3 ml/min flow rate to extract the total lipids. The AACC standard method (5819) for acid hydrolysis was used as a reference method. We extracted 5-10 % more total lipids by SFE than by acid hydrolysis depending on the sample matrix.
L.M. Seitz and M.S. Ram.
Chemical information for classifying grain odors was obtained
by using dynamic headspace technology coupled with gas chromatography-mass
spectroscopy to determine volatiles in a set of 745 samples consisting
of corn, sorghum, soybeans, and wheat. Sensory data for each sample
was obtained from at least two panels. Previous processing of
the chemical and sensory data by multivariate analyses such as
principal component analysis and partial least squares methods
helped to determine what volatiles could be used to classify grain
odors. In this study, we used artificial neural network methods
to classify odors in the samples. Proper choice of samples and
use of optimized variables (compounds indicating off-odors), as
well as some preprocessing of raw data were necessary for training
the networks. Properly trained, networks could classify samples
into two odor categories (normal and off) or as many as five categories
(normal, sour, musty, smoke, and insect) from analysis of chemical
data concerning relative amounts of specific volatile compounds
purged from each sample. Networks trained to classify into several
specific categories could also identify mixed odors, i.e., having
both musty and sour odors, in some samples. Panelists had given
a consensus assignment of only a single odor with some samples,
but neural network results and the corresponding presence of odor-indicating
compounds suggested that more than one type of odor could have
been assigned and may have been detected by some individual
panelists.
M.S. Ram, F. Dowell, and L. Seitz.
Soaking wheat kernels in an NaOH solution causes the difference in color to be enhanced. Red wheat turns a darker red and white wheat turns straw yellow upon soaking in NaOH solution. To understand the chemistry of this reaction, chromatographic and spectroscopic studies have been initiated. Acidified extracts have been analyzed by HPLC. Chromatograms from red and white wheat showed minor differences and were compared to those from scabby and rain-bleached wheat which appeared to have lost some pigmentation. Vibrational spectroscopic studies (FT-IR, FT-Raman, and IR-microscopy) of whole wheat and wheat bran are being continued.
M.S. Ram, L.M. Seitz, and F.E. Dowell.
An accurate, rapid, and simple means of determining wheat color class has been developed to aid in distinguishing between hard red and hard white varieties. Research by other scientists has shown that soaking kernels in dilute NaOH accentuates color differences resulting in a clear, objective differentiation between color classes. We optimized the procedure so that genetic color class can be determined at elevators in about 10 min. The test requires minimal training, is safe, and should cost only pennies per sample. This project was funded by the Kansas Wheat Commission and administered through a CRADA with the Grain Industry Alliance. Perten Instruments (Springfield, IL) has developed a commercial test kit for use in field locations. This technology will help keep white wheat segregated for use in new export markets.
M.S. Ram, L.M. Seitz, and F.E. Dowell.
It is occasionally necessary to tag wheat kernels without altering
their appearance. Examples include tagging genetically modified
grain, or grain with specific attributes of interest that need
to be identity preserved. Tagging wheat kernels could be achieved
using a coating of UV or NIR fluorescent chemical in combination
with a binder, such as a lacquer or matte finish paint. Coatings
could be superior to chemical derivatives, which alter the physical
properties of surfaces. A number of chemicals were tested, but
were discarded because of high carcinogenicity. Suitable materials
were chosen and methods developed to suitably coat red and white
wheat. UV fluorescence and NIR spectroscopy were applied to the
kernels with and without coatings. Most recently, the invisible
coating was used to tag Karnal bunt kernels used during calibration
of high-speed sorting instruments.
F.E. Dowell, M. Pasikatan, T.N. Boratynski, R. Ykema, A.K. Dowdy, and R.T. Staten.
A high-speed, optical sorter was used to remove kernels infected with Karnal bunt from 1,800-g wheat samples. When the sorter removed about 8 % or more of the sample, the reject portion contained 100 % of the bunted kernels. Concentrating the bunted kernels in a smaller sample size will reduce sample inspection time and should reduce inspection errors. One high-speed sorter can process up to 8,800 kg/hr, thus, bunted kernels can be rapidly removed from samples or large lots. The instrument sorted each sample in less than 1 minute. This technology provides the wheat industry with a tool to rapidly inspect samples to aid in regulating Karnal bunt, and to remove bunt from seed wheat and wheat destined for food or feed use. This sorter is currently being used by APHIS as part of their routine Karnal bunt inspection procedures. The high-speed sorter also was used to remove red wheat from hard white wheat stock. The popularity of white wheat is increasing in the Midwest HRWW-growing area because of possible higher prices that may be realized through demands in Asian noodle markets. We have used the sorter to rapidly remove red wheat from early-generation white wheat breeder samples and to remove red wheat from large lots of commercial seed. Removing red seed from breeder samples reduces the amount of red wheat in subsequent generations, and removing red wheat from commercial seed helps insure the purity of harvested wheat.
F.E. Dowell, M. Pasikatan, E. Maghirang, and D. Wang.
Some wheat-kernel attributes are not detectable by bulk-measuring devices. For example, defects that may be present in a small percentage of kernels, such as internal insects or fungal damage, may be detected only if each kernel is analyzed. Also, some types of blending, such as mixing high- and low-protein lots, may be detectable only by single-kernel analysis. In addition, for some applications such as purifying seed stock, it is desirable to not only detect the undesirable characteristic but to then also remove it. Technology includes 1) a low-cost NIR system for detecting and sorting single kernels at a rate of about 1 kernel/sec; 2) a Vis-NIR system (SKCS 4170, Perten Instruments) for detecting single-kernel attributes, including single-kernel hardness measurements, at about 1 kernel/sec; and 3) high speed Vis-NIR systems, such as those produced by Satake and Sortex, capable of sensing and sorting single kernels at a rate of hundreds of kernels/sec.
M.C. Pasikatan and F.E. Dowell.
Sorting systems based on optical methods have the potential
to rapidly detect and physically remove seeds severely contaminated
by fungi, or infested internally by insect larvae or pupae. Thus,
the literature on sorting systems based on optical methods for
detecting and sorting seeds with these attributes was reviewed.
Sorting indices based on wavelengths useful for detecting these
attributes were emphasized. Surface characteristics of seeds,
like discoloration caused by fungi, are generally detectable in
the visible range of the electromagnetic spectrum, whereas internal
attributes are detectable in the near-infrared range. The spectral
differences between sound and infested seeds are usually subtle,
but full-spectrum and two-wavelength classification models have
succeeded in detecting and classifying seeds based on these attributes.
For high sorting accuracies, wavelength identification and proper
selection of a sorting criterion are important. Chitin, ergosterol,
or hydrolysis of triglycerides have been identified as indicators
of seed fungal contamination whereas chitin, protein, phenolic
compounds, or changes in starch have been useful indicators of
internal insects in
seeds.
D. Wang, F.E. Dowell, and R. Dempster.
The content of dark hard vitreous (DHV) kernels in HRSW is
an important grading factor that is associated with protein content,
kernel hardness, milling properties, and cooking quality. The
current visual method of determining DHV and nonDHV (NDHV) wheat
kernels is time consuming, tedious, and subject to large errors.
Our objective was to classify DHV and NDHV wheat kernels including
kernels that are checked, cracked, sprouted, or bleached by using
near-infrared (NIR) spectroscopy. Spectra from single DHV and
NDHV kernels were collected using a diode-array NIR spectrometer.
The dorsal and crease sides of the kernels were viewed. Three
wavelength regions, 500-750 nm, 750-1,700 nm, and 500-1,700 nm,
were compared. Spectra were analyzed by using partial least squares
(PLS) regression. Results show that the major contributors to
classifying DHV and NDHV kernels are protein content, kernel hardness,
starch content, kernel color, and a scattering effect on the absorption
spectrum. Bleached kernels were the most difficult to classify.
The sample set with bleached kernels yielded lower classification
accuracies of 91.1-97.1 % compared to 97.5-100 % for the sample
set without bleached kernels. More than 75 % of misclassified
kernels were bleached. Classification models that included the
dorsal side gave the highest classification accuracies (99.6-100
%) for the testing sample set. Wavelengths in both the visible
and NIR regions or the NIR region alone yielded better classification
accuracies than these in the visible region only.
D. Wang, F.E. Dowell, and D.S. Chung.
Heat damage is a serious problem frequently associated with wet harvests because of improper storage of damp grain or artificial drying at high temperature. Heat damage causes protein denaturation and reduces processing quality. The current visual method for assessing heat damage is subjective and based on color change. The denatured protein related to heat damage does not always cause change in kernel color. NIR spectroscopy is possible method to measure both physical (color) and chemical (protein denaturation) changes. A diode-NIR spectrometer, which measured reflectance spectra (Log(1/R)) from 400 to 1,700 nm, was used to differentiate single kernels of heat-damaged and undamaged wheats. Partial least squares (PLS) regression was used to develop classification models with three wavelength regions (400-750 nm, 400-1,700 nm, and 750-1,700 nm). Results showed that the major contributor to the spectral characteristics of heat-damaged kernels is protein denaturation, which changes the patterns of molecular absorption and shifts the peaks of the absorption spectrum. For PLS models, the highest classification accuracy of 100 % for both calibration and testing sample sets was obtained from the NIR wavelength region of 750-1,700 nm. The visible wavelength region (400-750 nm) gave the lowest classification accuracy.
M.E. Casada.
Moisture adsorption rates for stored grains are important for accurate modeling of drying and storage. Wheat and barley samples at initial moisture contents typical of grain storage were exposed to several levels of higher humidity at two temperatures to measure adsorption rates. The best fit to the data was achieved with the Page and cellular diffusion equations. The adsorption rates were lower than those of comparable desorption tests. The adsorption rates for barley were lower than for wheat, due to lower diffusion coefficients for the barley endosperm and germ as compared to wheat.
S.R. Bean and T.C. Pearson.
Grain Quality and Structure Research Unit. Dr. Scott
Bean joined the Grain Quality and Structure Research Unit in
October as a Research Chemist. Dr. Bean received his M.S. and
Ph.D. degrees in Grain Science from the Department of Grain Science
and Industry, Kansas State University. Dr. Bean has his expertise
in cereal biochemistry and analytical methods for characterizing
cereal proteins using instruments including capillary electrophoresis
and liquid chromatography. In addition, Dr. Bean has experience
in studying structure-function relationships of cereal biomolecules
that relate to end-product quality. As lead scientist of the sorghum
project, his responsibilities will include the biochemical characterization
of grain sorghum for both human and feed uses, cultivar identification,
their relationships to functional and nutritional quality, and
providing information on quality biochemical determinants for
sorghum breeders to improve lines suitable for traditional and
novel uses. This is a new project created in response to efforts
by the National Grain Sorghum Producers to enhance sorghum value
in food and nonfood uses.
Engineering Research Unit. Dr. Tom Pearson joined the Engineering Research Unit in July to fill the position vacated by Dr. Inna Zayas. Dr. Pearson received his B.S. in mechanical engineering from California State University in Fresno, and his M.S. and Ph.D. in Agricultural Engineering from UC Davis. Dr. Pearson has experience as a project engineer/senior research and development engineer with Wiebe Conveyance, Hollister, CA, and as a lead scientist at the Western Regional Research Center, Albany, CA. He has expertise in developing high-speed detection and sorting systems, machine vision, NIR, acoustics, and electronic circuitry. Dr. Pearson's responsibilities at the GMPRC will include developing sensing methods, techniques, equipment, and instrumentation for automating quality assessments of grain and grain products.
C.M. Rosell.
The Grain Science and Industry Department (Dr. Paul A. Seib), Kansas State University, and the USDA-ARS-GMPRC-Grain Quality and Structure Research Unit (Dr. George L. Lookhart) co-hosted, during the summer of 2001, Dr. Cristina Molina Rosell from the Laboratorio de Cereales, Instituto de Agroquímica y Tecnología de Alimentos in Valencia, Spain. Dr. Molina came to this laboratory to learn capillary electrophoresis in order to study the effects of insects and enzymes on wheat storage proteins.