葡萄半不育花粉粒形态和细胞学观察及相关基因表达分析
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国家948项目[2011-G(28)];国家科技支撑项目(2013BAD20D05);江苏省三新工程项目(SXGC(2014)004);江苏省科技厅项目(SBE2014030811)


Morphological,Cytological Observation of Pollen Grains and Expression of Related Genes in Semi-sterile Grapes
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    摘要:

    以葡萄品种‘魏可’(可育)及其自交后代单株‘魏可实生-3’和‘魏可实生-12’以及品种‘钟山红’(雄性不育)为材料,对花器官特征、花粉萌发率、花粉粒形态以及小孢子发育过程的细胞学特征进行观察,并对绒毡层发育相关基因(DYT1、TDF1和MYB4)的表达进行分析,以初步鉴定葡萄半不育单株的不育特征,揭示花粉败育与基因调控的关系。结果显示:(1) ‘魏可实生-3’和‘魏可实生-12’ 的花粉萌发率均在10%左右,属于半不育花粉;其雄蕊主要特征为花丝卷曲,花丝短于花柱,花药内花粉量少,花药中既有正常长球形花粉粒也有异常近球形花粉粒。(2)‘魏可实生-3’、‘魏可实生-12’没有明显的药室壁结构,小孢子母细胞稀少、疏松;‘魏可实生-3’双核期绒毡层未完全降解,绒毡层细胞中的营养物质滞留没有及时供应小孢子的发育,致后期部分小孢子败育;‘魏可实生-12’绒毡层细胞在单核早期就大部分降解而导致营养供应异常,使部分小孢子败育。(3)在减数分裂期,‘魏可’的绒毡层发育相关基因DYT1和TDF1的表达均最高,MYB4最低,‘钟山红’的3个基因的表达与‘魏可’相反,‘魏可实生-3’和‘魏可实生-12’的表达水平均在‘魏可’与‘钟山红’之间;单核早期以后,3个基因在‘魏可’的表达一直均较低,钟山红’的DYT1基因表达一直最高,其余2个基因均在最低水平,而‘魏可实生-3’的TDF1、MYB4基因在单核早期表达量最高,‘魏可实生-12’ 的TDF1在双核期表达量最高。研究表明,‘魏可实生-3’的花粉败育发生在单核晚期至双核期,‘魏可实生-12’的花粉败育发生在单核早期;部分药室绒毡层的异常降解,小孢子发育营养供应不均衡可能是导致其花粉半不育发生的原因;‘魏可实生-3’的绒毡层延迟降解与减数分裂期DYT1基因的表达量不足、单核期TDF1基因的过量表达以及MYB4基因在单核期的异常表达有关,而‘魏可实生-12’的绒毡层提前降解与减数分裂期DYT1基因的表达量不足而MYB4基因过量表达,以及双核期TDF1基因过量表达有关。

    Abstract:

    ‘Wink’ (fertile),‘Zhongshanhong’ (male sterile),‘Wink seedling-3’,‘Wink seedling-12’ were examined.The flower characteristics,pollen viability,pollen shape and the cytological features of microspores were observed,DYT1,TDF1 and MYB4 genes that was related to the tapetum development were analyzed.The semi-sterile characteristics of the ‘Wink seedling-3’ and ‘Wink seedling-12’ were identified preliminarily,and the pattern of the gene regulation of pollen abortion was studied.The results indicated that:(1)the main features of two ‘Wink seedling’ grapes:filaments curl and shorter than the style,few pollen in the anther,the pollen viability was only about 10%.There were two kinds of pollen grains in anthers:normal prolate spheroidal pollen grains and approximately spheroidal pollen grains.(2)The anther wall structure was not obvious,and the microspore mother cells were few and loose in the ‘Wink seedling’ grapes.The tapetum of ‘Wink seedling-3’ did not degrade completely in the binuclear stage,the nutrients of tapetum could not supply for the development of spores timely,which caused the abortion of microspores in the late developmental stage.The tapetum of ‘Wink seedling-12’mostly disintegrated at the early mononuclear microspore,which caused the abnormal supply of nutrient and the abortion of microspores.(3)In the meiosis stage,DYT1 and TDF1 expression in ‘Wink’ were the highest,MYB4 was the lowest;These three gene expression in ‘Zhongshanhong’ were contrary to ‘Wink’,and the levels of two ‘Wink seedling’ grapes laid between ‘Wink’ and ‘Zhongshanhong’.After the mononuclear microspore stage,three gene expression in ‘Wink’ were always low,DYT1 expression levels were the highest,TDF1 and MYB4 were the lowest;TDF1 and MYB4 expression levels in ‘Wink seedling-3’ were highest in the mononuclear microspore stage,and TDF1 expression in ‘Wink seedling-12’ was highest in the binuclear stage.The study showed that ‘Wink seedling-3’abnormal tapetum degradation was probably related to the DYT1 insufficient expression in the meiosis stage,the TDF1 overexpression in the mononuclear microspore stage,and MYB4 unusual expression in the mononuclear microspore stage.‘Wink seedling-12’abnormal tapetum degradationwas probably related to the DYT1 insufficient expression and MYB4 overexpression in the meiosis stage,and the TDF1 overexpression in the binuclear stage.

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余晓娟,袁月,王继源,等.葡萄半不育花粉粒形态和细胞学观察及相关基因表达分析[J].西北植物学报,2015,35(7):1317-1325

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  • 在线发布日期: 2015-08-29
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