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GENETIC STOCKS

Registration of Hard Kernel Puroindoline Allele Near-Isogenic Line Hexaploid Wheat Genetic Stocks

^a USDA-ARS Western Wheat Quality Lab., E-202 Food Sci. & Human Nutrition Facility East, P.O. Box 646394, Washington State University, Pullman, WA 99164-6394
^b Dep. Food Sci. & Human Nutrition, Washington State University, Pullman, WA 99164-6394, assigned to the Western Wheat Quality Lab. Mention of trademark or proprietary products does not constitute a guarantee or warranty of a product by the U.S. Department of Agriculture and does not imply its approval to the exclusion of other products that may also be suitable

^* Corresponding author (morrisc{at}wsu.edu).

Abbreviations: NIL, near-isogenic line • SKCS, Single Kernel Characterization System • SWS, soft white spring

Seven puroindoline allele near-isogenic line (NIL) hexaploid wheat (Triticum aestivum L.) genetic stocks (GS-157–GS-163; PI 644080–PI 644086) were developed by Dr. Craig F. Morris at the USDA-ARS Western Wheat Quality Laboratory, Pullman, Washington. Since they incorporate the first seven known puroindoline mutations/alleles, they all express hard kernel texture. These genetic stocks were released by the USDA-ARS in February of 2007 because of the utility of such NILs in researching the effects of these puroindoline alleles on kernel texture and wheat grain end-use quality. Furthermore, these NILs, being developed in the ‘Alpowa’ (PI 566595) soft white spring (SWS) wheat cultivar background, will have direct benefit to the development of hard spring wheat cultivars in the Pacific Northwest and elsewhere. Alpowa has been the leading SWS wheat cultivar in Washington since 1997 (U. S. Dep. Agric. National Agric. Statistics Service).

These NILs (Table 1 ) were developed by selecting donor parents that possessed a unique puroindoline a and puroindoline b gene haplotype. Varieties with ‘functional’ or ‘wild-type’ puroindoline sequences (Pina-D1a/Pinb-D1a) have soft kernel texture while varieties with a mutation in puroindoline a or b have hard kernel texture. The tightly linked puroindoline genes represent the molecular-genetic basis of what is known as the Hardness locus on chromosome 5DS (Morris, 2002). Each donor parent was crossed as male to the soft white spring wheat cultivar Alpowa; F₁ seeds were harvested and planted; plants were allowed to self; F₂ seeds were harvested and planted; and plants were allowed to self. The F₃ seeds from individual F₂ plants were subjected to a progeny phenotypic screening for kernel texture using the Perten Instruments (Springfield, IL) Single Kernel Characterization System (SKCS) 4100 (Approved Method 55–31; AACC International, 2000). The F₂ plants were thus identified as being homozygous hard (ha/ha), homozygous soft (Ha/Ha) or heterozygous and segregating (1:2:1 Ha/Ha:Ha/ha:ha/ha) for puroindoline allele at the Hardness locus. A homozygous hard plant was selected for backcrossing using Alpowa as the recurrent male parent and the aforementioned process was repeated. In addition to hard kernel texture, at each cycle the progeny were selected for the morphological and developmental characteristics of Alpowa. In all, seven backcrosses were conducted such that the general pedigree of each NIL is: Alpowa/donor parent//7*Alpowa.

For the first of the NILs developed (Pina-D1a/Pinb-D1b), BC₇F₂-derived F₃ seed (BC₇F₂:F₃) was used to sow 1 m long single "plant" rows at the WSU Dep. of Plant Pathology Whitlow Farm, Pullman, WA in 2005. Each row was harvested separately, evaluated for kernel texture, and the puroindoline haplotype was confirmed via PCR and DNA gene sequencing (Morris, 2002). The BC₇F₂:F₄ seed from 10 selected plots was bulked and advanced; the present release is comprised of BC₇F₂:F₅ grain.

The remaining six NILs were obtained by the following, similar procedure: BC₇F₂:F₃ seed from multiple, individual plants of each of the six pedigrees was sown as small 1.2 x 1.2 m 4-row plots at the WSU Spillman Agronomy Farm, Pullman, WA in 2006. Field plots were harvested separately, evaluated for kernel texture (SKCS), and the puroindoline haplotype was confirmed via PCR and DNA sequencing, as appropriate (Morris, 2002). The Pina-D1b allele was confirmed using PCR primers that span the 15,380-bp deletion in Pina and downstream sequence (Morris and Bhave, 2007); the present release of these NILs is comprised of BC₇F₂:F₄ grain.

Morphologically and developmentally, these NILs are indistinguishable from Alpowa. Further, they are expected to carry the Yr39 gene for high temperature adult plant resistance to stripe (yellow) rust (caused by Puccinia striiformis West. f. sp. tritici) from Alpowa (Lin and Chen, 2007). SKCS kernel texture data on the BC₇F₂:F₃ field plots (BC₇F₂:F₄ seed) are provided in Table 1; these values are typical of those encountered during the several cycles of backcrossing. PI 644081 was tested as ‘WQL9HDALP’ in the Washington State University Extension Uniform Cereal Variety Testing Program (http://variety.wsu.edu; verified 27 Nov. 2007) hard white spring wheat nurseries in 2006 where it produced the top overall single location yield of 8.7 Mg ha^–1 and was not significantly different from the top-yielding variety in six of the other 15 locations. It was also tested in the Western Regional Hard Spring Wheat Nurseries in 2006 where it had the best (lowest) rank sum of all varieties across the nine locations.

Small quantities of seeds are available on written request to the corresponding author. It is requested that appropriate recognition be made if these genetic stocks contribute to research or the development of a new breeding line or cultivar. Genetic material of this release has been deposited in the National Plant Germplasm System where it will be available for research purposes, including development and commercialization of new cultivars.

Acknowledgments

The authors express their sincere thanks for the assistance of Drs. Xianming Chen and Kim Garland-Campbell, Dan Dreesman, Chris Hoagland, Dave Wood, Lynn Little, Shelle Freston, Stacey Sykes, and the staff of the Western Wheat Quality Laboratory.

Footnotes

All rights reserved. No part of this periodical may be reproduced or transmitted in any form or by any means, electronic or mechanical, including photocopying, recording, or any information storage and retrieval system, without permission in writing from the publisher. Permission for printing and for reprinting the material contained herein has been obtained by the publisher.

Received for publication February 21, 2007.

References

• AACC International. 2000. Approved Methods of the American Association of Cereal Chemists, 10th ed. AACC International, St. Paul, MN.
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• Donaldson, E., G.W. Bruehl, and G.L. Rubenthaler. 1991. Registration of Andrews wheat. Crop Sci. 31:1387.[Free Full Text]
• Lillemo, M., and C.F. Morris. 2000. A leucine to proline mutation in puroindoline b is frequently present in hard wheats from Northern Europe. Theor. Appl. Genet. 100:1100–1107.[CrossRef]
• Lin, F., and X.M. Chen. 2007. Genetics and molecular mapping of genes for race-specific all-stage resistance and non-race-specific high-temperature adult-plant resistance to stripe rust in spring wheat cultivar Alpowa. Theor. Appl. Genet. 114:1277–1287.[CrossRef][Medline]
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• Morris, C.F. 2002. Puroindolines: The molecular genetic basis of wheat grain hardness. Plant Mol. Biol. 48:633–647.[CrossRef][Medline]
• Morris, C.F., M. Lillemo, M.C. Simeone, M.J. Giroux, S.L. Babb, and K.K. Kidwell. 2001. Prevalence of puroindoline grain hardness genotypes among historically significant North American spring and winter wheats. Crop Sci. 41:218–228.[Abstract/Free Full Text]
• Morris, C.F., and Bhave, M. 2007. Reconciliation of D-genome puroindoline allele designations with current DNA sequence data. J. Cereal Sci. (in press) doi:10.1016/j.jcs.2007.09.012.
• Stewart, G. 1923. Sevier wheat. J. Amer. Soc. Agron. 15:385–392.

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