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远缘杂交是新优种质培育的主要途径,野生种质与栽培种进行远缘杂交在改良栽培品种以及培育新异种质中起着重要作用[1-3],如我国小麦育种专家李振声团队利用小麦(Triticum aestivum L.)与长穗偃麦草(Elytrigia elongata (Host) Nevski)杂交培育出抗病性强的‘小偃6号’等系列远缘杂交小麦品种,成功解决抗“小麦癌症”条锈病的世界育种难题[4],Moshe Feldman和Emest R.Sears也曾提出“未来的谷物改良寄希望于野生基因资源的利用”[5]。牡丹(Paeonia suffruticosa Andr.)为世界著名花卉,也是中国的一张名片,在中国影响之深远是其他花卉所不可企及的;但牡丹现有栽培品种中纯正花色少,黄色、橙色花缺乏,株型较低,大多对白粉病易感。长期以来,由于牡丹远缘杂交不亲和或亲和性差,牡丹育种多集中在品种群内进行常规杂交,遗传基础狭窄和多样性亲本资源的缺乏成为了限制牡丹育种取得突破的瓶颈。
栽培品种中,由杨山牡丹(P. ostii T.Hong et J.X.Zhang sp.nov.)演化出的‘凤丹’(P. ostii ‘Feng Dan’)植株高大、生长势强、花量大,生态适应性极广泛,是杂交育种最常用的亲本[6-7];牡丹野生种保康牡丹(Paeonia baokangensis Z. L. Dai et T. Hong sp. Nov.)[8],花桃红,色泽纯正、明亮,花期长,抗白粉病,是亟待开发的优良野生资源之一。解析牡丹‘凤丹’与保康牡丹远缘杂交的不亲和性机制,对于克服受精障碍,完善牡丹远缘杂交技术体系,培育花色纯正、抗白粉病的牡丹新品种具有重要意义。
植物远缘杂交研究发现,花粉萌发及花粉管生长情况、内源激素水平、保护酶活性及可溶性蛋白含量等是远缘杂交亲和性强弱的重要指标。郭朋辉等[9]在紫斑百合(Lilium nepalense D.Don)远缘杂交亲和性研究中发现,紫斑百合远缘杂交过程中多出现受精前障碍,胼胝质沉积,花粉管扭曲缠绕,阻碍伸长生长是其受精前障碍的主要表现;王保成等[10]在芸芥(Eruca sativa Mill)亲和性研究中发现,SOD、POD及CAT三种保护酶的活性变化与自交亲和基因的调控相关;杨晓玲等[11]在百合(Lilium brownie F.)亲和性研究中发现,高水平的IAA、GA3、ZR对花粉在柱头的识别黏附及萌发有促进作用;Kovaleva L 等[12]表明,ABA含量较高是导致矮牵牛(Petunia hybrida L.)授粉不亲和的原因;牡丹与芍药(Paeonia lactiflora Pall.)杂交后也显示相关保护酶活性及内源激素含量与授粉亲和性相关[13]。目前,有关牡丹种间杂交不亲和机制尚未见报道,基于此,本研究以牡丹‘凤丹’为母本,以野生保康牡丹为父本进行了种间杂交,探讨花粉萌发、花粉管生长、雌蕊保护酶活性、可溶性物质及内源激素含量变化与远缘杂交不亲和性的关系,以期为解析牡丹种间远缘杂交不亲和性机制以及完善牡丹远缘杂交技术体系提供理论及试验依据。
牡丹种间远缘杂交不亲和的细胞学与生理机制研究
Interspecific distant hybrid incompatibility cytology and its physiological mechanis in Paeonia suffruticosa Andr.
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摘要:
目的 明晰牡丹种间远缘杂交不亲和的细胞学及生理机制,为牡丹远缘杂交提供理论及试验基础。 方法 以牡丹‘凤丹’为母本,保康牡丹为父本,进行远缘杂交授粉亲和性研究,探讨授粉后雌蕊保护酶活性、可溶性物质含量及内源激素含量变化与花粉萌发、花粉管生长之间的联系。 结果 保康牡丹与‘凤丹’远缘杂交亲和性差的细胞学原因是花粉萌发慢,花粉管多有胼胝质沉积、扭曲缠绕严重,生长多无序并伴有分枝现象,只部分能穿过花柱到达子房。生理机制研究结果表明:授粉后,SOD活性快速升高有利于花粉柱头识别及花粉管生长;MDA含量快速增加不利于远缘杂交授粉花粉管的伸长生长;远缘杂交雌蕊较自交雌蕊可溶性蛋白及可溶性糖含量高,表现为柱头识别作用及雌蕊排异作用加强;自交雌蕊IAA含量显著高于杂交,高水平IAA、GA3、IPA有助于花粉萌发及花粉管生长;远缘杂交雌蕊ABA含量显著高于自交雌蕊,高水平ABA、JA对花粉管生长显示抑制作用,诱发胼胝质沉积,花粉管扭曲缠绕生长无序。 结论 ‘凤丹’与保康牡丹远缘杂交存在受精前障碍,其与雌蕊内部酶活性变化、内源激素含量的动态变化及可溶性物质含量密切相关。 Abstract:Objective To clarify the cytological and physiological mechanism of the incompatibility of distant hybridization between peony species, and provide theoretical and experimental basis for distant hybridization of peony. Method Using P. ostii ‘Feng Dan’ as the female parent and Paeonia baokangensis Z. L. Dai et T. Hong sp. Nov. as the male parent, the compatibility of distant hybridization and pollination was studied for exploring the relationship between the changes of pistils protect enzyme activity, soluble substance content and endogenous hormone content after pollination, pollens germination and pollen tubes growth. Result The results showed that the cytology of Paeonia baokangensis Z. L. Dai et T. Hong sp. Nov. and P. ostii ‘Feng Dan’ distant hybridization was poor as a result of slow pollens germination, corpus callosum deposition in pollen tubes, severe twisting and entanglement, disordered growth and branching, and only part of the pollens could pass through the style to the ovary. After pollination, the rapid increase of SOD activity was beneficial to pollens stigma recognition and pollen tubes growth; The rapid increase of MDA content was not conducive to the elongation and growth of pollen tubes in distant hybridization; Distant hybrid pistils had higher soluble protein and soluble sugar content than selfed pistils, which is manifested by enhanced stigma recognition and pistils rejection; The content of IAA in selfed pistils was significantly higher than that of distant hybridization, high levels of IAA, GA3 and IPA were helpful for pollens germination and pollen tubes growth; The ABA content of distant hybrid pistils was significantly higher than that of selfed pistils, high levels of ABA and JA showed an inhibitory effect on the growth of pollen tubes, induced the deposition of callosum, and the pollen tubes twisted and twisted to grow disorderly. Conclusion Pre-fertilization barriers exist in distant hybridization of P. ostii ‘Feng Dan’ and Paeonia baokangensis, which is closely related to the changes of enzyme activity in the pistil, the dynamic changes of endogenous hormone content and the content of soluble substances. -
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