‘无籽’瓯柑小孢子母细胞减数分裂特性基因RAD51和MS1的表达差异分析

发布时间：2017-12-27 08:45

本文关键词：‘无籽’瓯柑小孢子母细胞减数分裂特性基因RAD51和MS1的表达差异分析　出处：《浙江农林大学学报》2016年06期 　论文类型：期刊论文

【摘要】：‘无籽’瓯柑Citrus suavissima‘Seedless’是野生型瓯柑Citrus suavissima的芽变,其花粉完全败育,败育起始于小孢子母细胞减数分裂形成四分体时期。研究‘无籽’瓯柑小孢子母细胞减数分裂异常的分子机制,有利于深入认识果树芽变的发生机制。随机选择"无籽"瓯柑和瓯柑各3株,根据花蕾直径采集花粉母细胞时期(Ⅰ),四分体时期(Ⅱ),单核花粉粒时期(Ⅲ),双核花粉粒时期(Ⅳ)及花粉粒成熟期(Ⅴ)共5个时期的花蕾并分离花药,利用实时荧光定量聚合酶链式反应(q RT-PCR)分析减数分裂特性基因RAD51和MS1的相对表达量,3次重复。通过SPSS 16.0的最小显著差法(LSD)在0.01水平上进行各样品间的相对表达量差异分析。结果表明:RAD51和MS1基因的表达高峰均集中在第Ⅰ时期和第Ⅱ时期,且此时花药中的相对表达量均极显著(P0.01)高于花蕾。第Ⅰ时期花药中,‘无籽’瓯柑RAD51的相对表达量为瓯柑的3.7倍,MS1可达300倍;但同时期花蕾中,RAD51的相对表达量仅1.1倍,MS1为140.0倍。‘无籽’瓯柑RAD51基因在第Ⅰ时期和第Ⅱ时期的相对表达量极显著(P0.01)高于瓯柑,说明‘无籽’瓯柑小孢子母细胞和四分体时期的细胞内均存在DNA受损现象;‘无籽’瓯柑MS1基因在第Ⅰ时期的相对表达量极显著(P0.01)高于瓯柑,但第Ⅱ时期极显著(P0.01)低于瓯柑,推测‘无籽’瓯柑败育花粉的形成与MS1基因表达紊乱引起油脂分泌、转运异常密切相关。
[Abstract]:'seedless Ougan' Citrus suavissima 'Seedless' is a wild type citrus suavissima Citrus suavissima mutant, the pollen abortion abortion completely, starting at meiosis of microspore mother cells was divided into four periods. "Study on the molecular mechanism of Seedless Ougan 'abnormal division of microspore mother cell meiosis, is conducive to in-depth understanding of the mechanism of fruit sport. Randomly selected "Seedless" mandarin orange and Mandarin Orange 3 strains, according to the collection of bud diameter pollen mother cell stage (I), (II) the four tetrad stage, mononuclear pollen stage (III), (IV) binucleate pollen stage and pollen mature stage (V) a total of 5 times bud and anther from, using real-time fluorescence quantitative polymerase chain reaction (Q RT-PCR) expression analysis of meiotic identity genes RAD51 and MS1, repeated 3 times. The difference in the relative expression of all kinds of products was analyzed at the 0.01 level by the minimum significant difference method (LSD) of SPSS 16. The results showed that the peak of RAD51 and MS1 gene expression was concentrated in stage I and II, and the relative expression level of anther was very significant (P0.01) higher than that of buds. The first period of anther, 'seedless Ougan' the relative expression of RAD51 was 3.7 times of mandarinorange, MS1 up to 300 times; but in the same period in the flower buds, the relative expression of RAD51 is only 1.1 times, 140 times MS1. 'seedless Ougan' RAD51 gene in the first period and the second period of relative expression was significantly higher than that of mandarin orange (P0.01), that "there were damaged DNA seedless Ougan 'microsporocyte tetrad stage and four cells;' seedless Ougan 'MS1 gene in 1 during the period of relative expression was significantly higher than that of mandarin orange (P0.01), but the second period was significantly lower (P0.01) Ougan, that' seedless Ougan 'abortive formation and expression of MS1 gene in pollen fertility disorder caused by oil secretion, transport is closely related to the abnormal.
【作者单位】：浙江农林大学亚热带森林培育国家重点实验室培育基地;浙江农林大学浙江省农产品品质改良技术研究重点实验室;
【基金】：国家自然科学基金资助项目(31000897) 浙江省自然科学基金资助项目(Y3090494) 浙江省团队科技特派员服务计划项目(浙科发农〔2013〕215-122) 浙江省大学生科技创新活动计划项目(2016R412003)
【分类号】：S666.1
【正文快照】： (1.The Nurturing Station for the State Key Laboratory of Subtropical Silviculture,Zhejiang AF University,Lin’an311300,Zhejiang,China;2.The Key Laboratory for Quality Improvement of Agriculture Products of Zhejiang Province,Zhejiang AF University,Lin’

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