Transcriptomic Microarray Analysis Revealed Acetyl-CoA Metabolic Pathway Defect in Male Sterile Line BNS in Wheat (Triticum aestivum)
FU Qing-Yun1, YANG Jing1, SUN Hai-Li1, SU Qing1,2, WEI Xiao1, RU Zhen-Gang1*, LI You-Yong1*
1 School of Life Science and Technology, Henan Institute of Science and Technology/Collaborative Innovation Center of Modern Biological Breeding, Xinxiang 453003, China; 2 College of Life Science, Henan Normal University, Xinxiang 453003, China
摘要BNS (Bai-nong sterility)是一个普通小麦(Triticum aestivum)的核型温敏雄性不育系,在杂交小麦研究中有重要价值。为揭示其不育生理机制,本研究采用 Affymetrix 小麦表达谱芯片检测 BNS 雄性不育 和雄性可育四分体时期、单核期花药及对应穗轴的转录组,分析差异探针谱生成的代谢网络。结果发现, BNS 可育花药上调表达探针基因生成的代谢网络中,形成乙酰辅酶 A 代谢子网络,完全不育花药中该代谢子网络缺陷,部分不育花药中缺少合成节点。乙酰辅酶 A 代谢子网络由乙酰辅酶 A 合成和代谢、酰基 辅酶 A 合成和代谢、硫酯合成和代谢 6 个节点组成,连接的代谢途径有激素代谢、蜡质代谢、氨基酸代谢、 ATP 合成、线粒体电子传递等,继而连接柱头、花药和花粉发育。乙酰辅酶 A 是细胞能量代谢的枢纽性物质,该代谢途径缺陷直接影响 ATP 等能量载体生成,从而影响小孢子发育。分别检测 BNS 不育株和可育株的花药、旗叶组织中 ATP 含量,发现不育花药中 ATP 含量比可育花药下降 10% 左右(P<0.01),不育旗叶中 ATP 含量比可育旗叶下降达 50% (P<0.01)。上述结果证实不育系在小孢子发育关键时期出现 ATP 供应短缺,推测乙酰辅酶 A 代谢缺陷是 BNS 雄性不育的重要生理机制。本研究为揭示 BNS 的不育发生 过程提供了重要线索,也为植物雄性不育能量供应短缺的形成提供了代谢依据。
Abstract：BNS (Bai-nong sterility) is a nuclear thermo-sensitive male sterile line of common wheat (Triticum aestivum), which plays an important role for the study of hybrid wheat. In order to explore the physiological mechanism of BNS sterility, Affymetrix wheat microarray was used to detect the transcriptomes of BNS male sterility and male fertile in anthers and spike-stalks in the developmental stage of tetrad and mononuclear, and analyzed metabolic networks generated by differential probe genes. The results found that an acetyl-CoA metabolic sub-network could be formed in the profiles of up-regulated genes in fertile anthers, but the sub- network was incompletely in partly sterile anthers, and it was not formed in sterile anthers. The acetyl-CoA sub-metabolic network consisted of 6 nodes, which were acetyl-CoA synthesis and metabolism, acyl-CoA synthesis and metabolism, and thioester synthesis and metabolism, respectively. The nodes of sub-network of acetyl-CoA first linked to that of hormone metabolism, waxy metabolism, amino acid metabolism and linked to ATP synthesis, mitochondrial electron transfer, and so on, then connected with stigma and anther development. Acetyl-CoA was a pivotal substance in cell energy metabolism, so the deficiency of this metabolic pathway directly affected the production of energy carriers such as ATP, thus affecting microspore development. ATP content in anther and flag leaf tissue of BNS sterile and fertile plants were detected, respectively. The results showed the content of ATP in the sterile anther decreased by about 10% compared with fertile anther (P<0.01), and the content of ATP in the sterile flag leaf decreased by 50% compared with fertile flag leaf (P<0.01). The results indicated that there was a shortage of ATP supply in the critical period of microspore development. Thus it was believed that the metabolic defect of acetyl-CoA was an important physiological mechanism for BNS male sterility. The mechanism provides an important clue for revealing the occurrence of BNS infertility.
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