Identification of microRNAs potentially involved in male sterility of Brassica campestris ssp. chinensis using microRNA array and quantitative RT-PCR assays.
Identification of microRNAs potentially involved in male sterility of Brassica campestris ssp. chinensis using microRNA array and quantitative RT-PCR assays.
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DOI：
10.2478/s11658-013-0097-9
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发表时间：
2013-09
影响因子：
8.3
通讯作者：
Cao J
中科院分区：
文献类型：
--
作者：
Jiang J;Jiang J;Yang Y;Cao J
MicroRNAs(MiRNAs)是一类新发现的非编码小RNA分子，可以负向调节基因的表达。据报道，许多miRNAs参与植物的生长、发育和逆境反应过程。然而，它们在开花植物有性繁殖机制中的作用仍不清楚。花粉发育是开花植物生活史上的一个重要过程，与作物种子的产量和品质密切相关。本研究旨在鉴定与花粉发育相关的miRNAs。以已知的植物miRNAs互补序列为探针，对甘蓝型油菜不育系(Bcajh97-01A)和可育系(Bcajh97-01B)的花序进行了基因芯片分析。中国草本植物‘矮交黄’核不育姊妹系(Bcajh97-01A/B)。结果表明，44个miRNAs在两个品系中有不同的表达。其中15个的转录水平发生了1.5倍以上的变化，其中9个在Bcajh97-01a不育系的花序中上调，6个下调。然后，我们重点研究了这15个miRNAs中的3个(miR158、MIR168和miR172)。通过计算方法预测了这3个miRNAs的13个家族成员，并预测了22个基因作为它们的候选靶基因。利用5‘端改良RACE方法，鉴定了MIR168的2个靶基因和miR172的5个靶基因。然后，用qRT-PCR方法验证了这3个miRNAs在花芽发育的5个时期的存在和表达模式。结果与基因芯片结果基本一致。因此，本研究可能为进一步探索可能在花粉发育过程中发挥作用的miRNA组提供有价值的线索。本文的补充材料可通过10.2478/s11658-0130097-9获得，授权用户可访问。
microRNAs (miRNAs) are a class of newly identified, noncoding, small RNA molecules that negatively regulate gene expression. Many miRNAs are reportedly involved in plant growth, development and stress response processes. However, their roles in the sexual reproduction mechanisms in flowering plants remain unknown. Pollen development is an important process in the life cycle of a flowering plant, and it is closely related to the yield and quality of crop seeds. This study aimed to identify miRNAs involved in pollen development. A microarray assay was conducted using the known complementary sequences of plant miRNAs as probes on inflorescences of a sterile male line (Bcajh97-01A) and a fertile male line (Bcajh97-01B) of the Brassica campestris ssp. chinensis cv. ‘Aijiaohuang’ genic male sterility sister line system (Bcajh97-01A/B). The results showed that 44 miRNAs were differently expressed in the two lines. Of these, 15 had over 1.5-fold changes in their transcript levels, with 9 upregulated and 6 downregulated miRNAs in inflorescences of ‘Bcajh97-01A’ sterile line plants. We then focused on 3 of these 15 miRNAs (miR158, miR168 and miR172). Through computational methods, 13 family members were predicted for these 3 miRNAs and 22 genes were predicted to be their candidate target genes. By using 5’ modified RACE, 2 target genes of miR168 and 5 target genes of miR172 were identified. Then, qRT-PCR was applied to verify the existence and expression patterns of the 3 miRNAs in the flower buds at five developmental stages. The results were generally consistent with those of the microarray. Thus, this study may give a valuable clue for further exploring the miRNA group that may function during pollen development. Supplementary material is available for this article at 10.2478/s11658-013-0097-9 and is accessible for authorized users.
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影响因子：
14.9
作者：
Dai X;Zhao PX
通讯作者：
Zhao PX
影响因子：
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作者：
Jones-Rhoades, MW;Bartel, DP
通讯作者：
Bartel, DP
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作者：
Katiyar-Agarwal S;Jin H
通讯作者：
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通讯作者：
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作者：
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通讯作者：
Van de Peer, Yves