花生栽培种和野生种Arachis sp.30119六倍体杂种的创制与鉴定

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中文摘要：[目的]花生野生种包含许多优异的抗病虫基因，是栽培种改良的重要基因资源库，将野生种染色质导入栽培种是花生远缘杂交研究的重要目的。野生种A。sp。30119对多种花生病害具有抗性，研究其与栽培种的杂种后代，为明确A。sp。30119的身份和未来能够利用其优异抗性提供依据。[方法]利用异源四倍体花生栽培品种白突 131 与二倍体野生种A。sp。30119杂交，通过胚拯救获得种间杂种 F1(W1212)，但 F1 不结实。为了揭示 W1212 不结实的原因并获得可育的后代，先利用根尖细胞有丝分裂中期和花粉母细胞染色体制片，观察其染色体数目和减数分裂染色体联会情况，再利用试管苗染色体加倍方法对 W1212 进行加倍处理，最终收获 4 个单粒荚果，将其中一个荚果的种子组织培养成完整植株并命名为Am1212。利用顺序GISH/FISH和SSR标记分析Am1212 的染色体组成，并观察调查其表型特征。最后，利用rDNA FISH随机分析 8 个Am1212 的F3 代植株的有丝分裂中期染色体数目，评价其染色体遗传稳定性。[结果]白突 131 与A。sp。30119的杂种一代W1212 花粉母细胞减数分裂中期Ⅰ的染色体平均构型为 1Ⅲ+6Ⅱ+15Ⅰ，其染色体不能正常配对，表现为高度不育，染色体加倍处理后W1212 下针枝条的花粉育性显著提高。白突 131、A。sp。30119和Am1212 的顺序GISH/FISH分析结果表明，A。sp。30119可能为A基因组二倍体野生种。Am1212 有 60 条染色体，包含白突 131 与A。sp。30119的全部染色体，证实其为六倍体杂种后代。但Am1212 自交F3 代有 37。5%的植株发生了染色体数目变异。通过分子标记筛选和表型调查，获得 17 个特异追踪A。sp。30119的显性和共显性标记，明确了Am1212 的遗传特性。[结论]创制了一个新的六倍体花生Am1212，该六倍体整合了野生种染色质，同时，Am1212 也拥有了小荚果等不利性状，且染色体遗传稳定性较差，因此，未来育种利用过程中，需要结合精准的染色体鉴定技术，打破不良连锁，获得具有补偿效应且包含有利性状的染色体变异材料。

外文标题：Development and Identification of an Interspecific Hexaploid Hybrid Between an A.hypogaea Cultivar and a Wild Species Arachis sp.30119 in Peanut

外文摘要：[Objective]Wild Arachis species containing many elite disease and insect resistant genes are an important gene pool for the improvement of cultivated peanut A.hypogaea L.,so the introduction of chromatin from wild species into cultivated peanut remains an important task for distant hybridization of peanut.Unfortunately,only a few wild species have been successfully involved in the development of interspecific hybrids,despite the existence of a huge amount of wild germplasm resource.The wild species A.sp.30119 contains multiple disease resistance.However,no interspecific hybrid between it and the cultivated peanut has been reported.[Method]After crossing allotetraploid peanut cultivar Baitu 131 with diploid wild species A.sp.30119 and followed by the embryo rescue,the interspecific hybrid F1(W1212)was obtained.However,interspecific hybrid F1 did not produce seed when selfed.In order to reveal the reason for infertility and continue the generation,the chromosome number of root tip cells was counted and chromosome pairing in pollen mother cells during meiosis of W1212 observed.The chromosome of W1212 was doubled through colchicine treatment during the in vitro culture W1212 plantlet.Finally,we harvested four one-seeded pods from the established W1212 plants;one of the pods undergone in vitro cultured and a number of plants were established from the culture and named as Am1212.The chromosome composition of Am1212 was analyzed by sequential GISH/FISH and SSR marker.Additionally,phenotypic characteristics of Am1212 were investigated.The metaphase chromosome numbers of 8 randomly selected F3 plants were analyzed by rDNA FISH to evaluate the genetic stability of Am1212.[Result]The average chromosome configuration during meiosis of pollen mother cells of W1212 was 1 Ⅲ+6Ⅱ+15Ⅰ,and the abnormalities in chromosome pairings lead to the high sterility of F1 plants.The pollen viability of peg-setting branches of W1212 was significantly improved after chromosome doubling treatment.Sequential GISH/FISH of Baitu 131,A.sp.30119 and Am1212 indicated that A.sp.30119 may be a diploid with A genome.Am1212 has 60 chromosomes consisting of all chromosomes of both Baitu 131 and A.sp.30119,which confirmed its nature of a hexaploid hybrid,but 37.5%F3 plants derived from Am1212 exhibited chromosome number variation.After conducting the screening of SSR molecular markers and phenotypic investigation,we obtained 17 dominant or co-dominant markers for specifically tracking wild species chromosomes and clarified the genetic characteristics of Am1212.[Conclusion]In this study,we successfully created a new hexaploid peanut Am1212 which incorporated A.sp.30119 chromatin.However,Am1212 exhibited instability in chromosome numbers and inherited unacceptable agronomic traits from wild species,such as small pods.Therefore,it is necessary to develop more accurate and efficient chromosome manipulation techniques to minimize the unfavorable gene linkage and generate alien chromosomal lines that possess compensatory effects and favorable traits for future breeding purposes.

外文关键词：

peanutwild specieshexaploidoligonucleotide fluorescence in situ hybridizationmolecular markers

作者：

刘华、曾凡佩、王倩、陈国权、苗利娟、秦利、韩锁义、董文召、杜培、张新友

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作者单位：

河南省农业科学院河南省作物分子育种研究院/神农种业实验室/农业农村部黄淮海油料作物重点实验室/河南省油料作物遗传改良重点实验室,郑州 450002

郑州大学农学院,郑州 450001

郑州大学生命科学学院,郑州 450001

关键词：

花生野生种六倍体寡核苷酸荧光原位杂交分子标记

基金：

河南省农科院优秀青年基金国家自然科学基金河南省科技研发计划联合基金现代农业产业技术体系建设项目河南省现代农业产业技术体系建设项目河南省农科院基础科研项目

项目编号：

2022YQ1632272153232301420025CARS-13S2012-052023JC03

出版年：

2024

DOI：

10.3864/j.issn.0578-1752.2024.10.003

中国农业科学

中国农业科学院

中国农业科学

CSTPCD北大核心

影响因子：1.899

ISSN：0578-1752

年,卷(期)：2024.57(10)

参考文献量33