Identification of responsive miRNAs involved in combination stresses of phosphate starvation and salt stress in soybean root
Identification of responsive miRNAs involved in combination stresses of phosphate starvation and salt stress in soybean root
复制标题
大豆根系磷酸盐饥饿和盐胁迫联合胁迫中响应性 miRNA 的鉴定
DOI：
10.1016/j.envexpbot.2019.103823
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发表时间：
2019-11-01
影响因子：
5.7
通讯作者：
Yu, De-Yue
中科院分区：
文献类型：
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作者：
Ning, Li-Hua;Du, Wen-Kai;Yu, De-Yue
盐胁迫和磷饥饿严重影响植物的生长发育。在许多植物中，已经报道了参与PI饥饿或盐胁迫的单一胁迫的microRNAs。然而，有关PI饥饿和盐胁迫响应的microRNAs及其靶基因的联合胁迫研究尚未见报道。在本研究中，我们利用Illumina HiSeqTM2000测序技术，在4种不同处理下构建了7个小RNA文库，其中包括CK、盐胁迫(SR1、SR5)、PI饥饿(LPR1、LPR5)和盐胁迫/PI饥饿双重胁迫(DR1、DR5)。共鉴定出5285个miRNAs，其中3616个为保守miRNAs，1669个为新miRNAs。其中，GMA-miR1691-3p、GMA-miR5036、GMA-miR862a和GMA-398a/b共检测到21个miRNAs，它们对PI饥饿和盐胁迫的单一胁迫和双重胁迫有反应。已鉴定的miRNAs的靶基因如乙烯反应转录因子4、蛋白磷酸酶2C和超氧化物歧化酶[铜-锌]，被分为不同的生物过程，包括代谢过程、信号转导。RT-qPCR验证了几个已鉴定的miRNAs的表达模式，RT-qPCR的结果与IlllightaHiSeq的结果一致。测定了不同处理下大豆根、叶中Na+、K+、PO43-的含量。我们的研究结果证实了差异表达的miRNAs响应PI饥饿和盐胁迫的复合胁迫，为提高大豆对盐胁迫和PI饥饿的抗性提供了必要的信息。
Salt stress and phosphate (Pi) starvation severely affect plants growth and development. MicroRNAs involved in single stress of Pi starvation or salt stress have been reported in many plant species. However, the combination stresses of Pi starvation and salt stress responsive microRNAs and their target genes have not been reported. In the, study, we generated 7 small RNA libraries under 4 different treatments with three independent biological replicates: CK, salt stress (SR1, SR5), Pi starvation (LPR1, LPR5), and the double stresses of salt stress/Pi starvation (DR1, DR5) in soybean root by using Illumina HiSeqTM2000 sequencing technique. Total of 5285 miRNAs including 3616 conserved and 1669 novel miRNAs were identified. Among them, 21 miRNAs including gma-miR1691-3p, gma-miR5036, gma-miR862a, and gma-398a/b, were detected to respond to the single stress and the double stresses of Pi starvation and salt stress. The target genes for the identified miRNAs such as ethylene-responsive transcription factor 4, protein phosphatase 2C, and superoxide dismutase [Cu-Zn], were classified into different biological processes including metabolic process, signaling transduction. The expression patterns of several identified miRNAs were validated by RT-qPCR and the results of RT-qPCR were consistent with the result of IlluminaHiSeq. The Na+, K+, and PO43- contents under different treatments were detected in soybean roots and leaves. Our findings identified the differentially expressed miRNAs in response to the combination stresses of Pi starvation and salt stress, which providing the essential information for enhancing the resistance to salt stress and Pi starvation in soybean.