小麦-长穗偃麦草7E抗赤霉病易位系培育

doi:10.3864/j.issn.0578-1752.2016.18.002

摘要/Abstract

摘要： 【目的】将二倍体长穗偃麦草7E染色体导入主栽小麦背景，培育小麦-长穗偃麦草7E染色体抗赤霉病易位系，为小麦抗赤霉病遗传改良利用外源优良基因提供新种质。【方法】利用中国春-长穗偃麦草7E代换系DS7E（7B）与扬麦16杂交的F₂种子进行⁶⁰Co辐射（30 000 rad）和种植，表现型选择收获存活M₁植株的种子，从M₂连续通过表型农艺性状选择、单花滴注法进行赤霉病抗性鉴定和长穗偃麦草7E染色体或染色体臂特异分子标记PCR扩增筛选，最后在M₄代对中选材料以长穗偃麦草基因组DNA为探针进行基因组原位杂交（genomic in situ hybridization，GISH）证实。【结果】M₁选择了赤霉病发病率不同的13个单株进行繁殖。利用前期开发的长穗偃麦草7E染色体和7EL、7ES特异标记检测13株的后代M₂单株，获得含有长穗偃麦草7EL片段7株和7ES片段14株；对21株M₂衍生的222个M₃植株进行特异标记检测，共选择含有长穗偃麦草7EL片段13株和7ES片段3株；利用来自12株M₃的后代（M₄）进行GISH，共9株M₃的后代具有小麦-长穗偃麦草易位染色体，体细胞染色体2n=42。2株M₃的后代显示附加2条长穗偃麦草染色体短臂，体细胞染色体2n=44。连续多年多途径的筛选，获得4份材料，3份材料均为长穗偃麦草7E染色体长臂易位系，命名为TW-7EL1、TW-7EL2和TW-7EL3。1份为7E染色体短臂附加系，命名为W-DA7ES，最后所获得的材料是源自M₁代2个单株。连续3年赤霉病抗性鉴定结果表明长穗偃麦草7EL易位系抗性高，发病率明显低于中国春和扬麦16，与苏麦3号相当，而7ES附加系的赤霉病抗性明显较低，发病率明显高于7EL易位系。【结论】通过赤霉病抗性鉴定、染色体特异分子标记筛选和GISH证实相结合培育了小麦-长穗偃麦草7EL抗赤霉病易位系，长穗偃麦草易位片段鉴定快速和准确。二倍体长穗偃麦草7E染色体长臂中含有抗小麦赤霉病的基因。

关键词: 小麦, 长穗偃麦草, 赤霉病, 易位系

Abstract: 【Objective】The objective of this study was to transfer the chromosome 7E of Thinopyrum elongatum into cultivated common wheat (Triticum aestivam L.) to develop translocation lines resistant to Fusarium Head Blight (FHB) and thereby to provide new germplasm for improving FHB resistance in common wheat.【Method】The F₂ seeds from the cross between Yangmai16 and DS7E(7B) were radiated using ⁶⁰Co (at 30 000 rad), DS7E(7B) being a substitution line in which the chromosome 7B of Chinese Spring common wheat was substituted with the chromosome 7E of Th. elongatum. The survived M₁plants were harvested, after visual selection for agronomic traits. The M₂ to M₄ populations were selected for agronomic traits, FHB resistance under single drop injection with Fusarium graminearum, and molecular markers specific to the chromosome and chromosomal arm of 7E for Th. elongatum, followed by cytological confirmation for the presence of 7E chromosomes using Th. elongatum genomic DNA as probe by genomic in situ hybridization (GISH). 【Result】Thirteen M₁ plants with varying degrees of FHB resistance were selected, and the corresponding M₂ plants were examined for the presence of previously developed molecular markers specific to chromosome 7E . Seven plants were found to carry the long arm of 7E and 14 were found to carry the short arm of 7E. After selfing, 13 plants carrying markers specific to the long arm of 7E chromosome and 3 plants carrying markers specific to the short arm of 7E chromosome were identified out of 223 M₃ plants. GISH analysis was conducted for the progenies (M₄) derived from 12 M₃ plants and it was found that the progenies from nine of the M₃ lines were wheat-Th. elongatum translocation lines (2n=42), and those from two other M₃ plants were chromosome addition lines with the short arm of 7E (2n=44). Continued selection led to the development of three translocation lines carrying the long arm of 7E, which were named as TW-7EL1, TW-7EL2 and TW-7EL3, respectively. A fourth line was a chromosome addition line with the short arm of 7E and was named as W-DA7ES. These four lines were derived from two different M₁ plants. Evaluation of FHB resistance indicated that the translation lines were similar to Sumai 3 in FHB resistance, better than Chinese Spring and Yangmai 16, while the addition line was considerably poorer in FHB resistance.【Conclusion】Translocation lines with chromosome 7EL that are resistant to FHB were developed effectively and accurately by joint use of phenotypic selection, screening for chromosome-specific molecular markers to 7E, and genomic in situ hybridization. The chromosome 7EL of Th.elongatum carries FHB-resistant genes.

Key words: wheat, Thinopyrum elongatum, fusarium head blight, translocation lines

张璐璐，陈士强，李海凤，刘慧萍，戴毅，高勇，陈建民. 小麦-长穗偃麦草7E抗赤霉病易位系培育[J]. 中国农业科学, 2016, 49(18): 3477-3488.

ZHANG Lu-lu, CHEN Shi-qiang, LI Hai-feng, LIU Hui-ping, DAI Yi, GAO Yong, CHEN Jian-min. Development of Wheat -Thinopyrum elongatum Translocation Lines Resistant to Fusarium Head Blight[J]. Scientia Agricultura Sinica, 2016, 49(18): 3477-3488.

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