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南方型紫花苜蓿根系盐胁迫应答转录因子鉴定与分析 被引量:8
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作者 马进 郑钢 +1 位作者 裴翠明 张振亚 《浙江农林大学学报》 CAS CSCD 北大核心 2016年第2期201-208,共8页
转录因子可以调节众多下游基因的表达,在植物抗逆境中起重要的调节作用。为了解析转录因子在南方型紫花苜蓿适应盐胁迫环境的分子机制,以南方型紫花苜蓿Medicago sativa‘Millennium’为材料,以正常培养(WT_ck1)和氯化钠(盐)胁迫(WT_N1... 转录因子可以调节众多下游基因的表达,在植物抗逆境中起重要的调节作用。为了解析转录因子在南方型紫花苜蓿适应盐胁迫环境的分子机制,以南方型紫花苜蓿Medicago sativa‘Millennium’为材料,以正常培养(WT_ck1)和氯化钠(盐)胁迫(WT_N1)条件下的2个样品根系进行转录组分析,鉴定紫花苜蓿根系盐胁迫应答转录因子基因。同时,随机挑选4个转录因子差异表达基因进行实时荧光定量q RT-PCR(3次重复),验证转录组测序技术(RNA-Seq)结果的可靠性。结果表明:紫花苜蓿根系在250 mmol·L-1氯化钠胁迫下72 h,共检测到31 907个基因表达量发生了改变,表达量差异达到2倍以上的基因共2 758个。其中,隶属于38个转录因子家族199个转录因子在盐胁迫下差异表达,上调表达104个,下调表达95个。在各转录因子家族中,盐胁迫应答基因数量最多的是MYB基因家族,其后分别是AP2-EREBP,b HLH,WRKY,NAC和GRAS基因家族,这暗示了紫花苜蓿根系对盐胁迫响应可能是多种转录因子家族共同参与的应答过程。q RT-PCR分析表明:4个随机选择的基因在胁迫前后的表达特点与表达谱测序结果一致。此外,Ms ERF-2b,Msb HLH,Msb ZIP,Ms GRAS,Ms NAC,Ms MGT-3a和Ms WRKY等转录因子被选为与盐胁迫应答相关的候选转录因子。该研究结果为阐明植物对盐胁迫的应答机制提供了新的线索。 展开更多
关键词 植物育种学 南方型紫花苜蓿 转录因子 根转录 盐胁迫
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Screening for differentially expressed genes in Anoectochilus roxburghii (Orchidaceae) during symbiosis with the mycorrhizal fungus Epulorhiza sp. 被引量:7
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作者 LI Biao TANG MingJuan +2 位作者 TANG Kun ZHAO LiFang GUO ShunXing 《Science China(Life Sciences)》 SCIE CAS 2012年第2期164-171,共8页
Mycorrhizal fungi promote the growth and development of plants, including medicinal plants. The mechanisms by which this growth promotion occurs are of theoretical interest and practical importance to agriculture. Her... Mycorrhizal fungi promote the growth and development of plants, including medicinal plants. The mechanisms by which this growth promotion occurs are of theoretical interest and practical importance to agriculture. Here, an endophytic fungus (AR-18) was isolated from roots of the orchid Anoectochilus roxburghii growing in the wild, and identified as Epulorhiza sp. Tissue-cultured seedlings of A. roxburghii were inoculated with AR-l 8 and co-cultured for 60 d. Endotrophic mycorrhiza formed and the growth of A. roxburghii was markedly promoted by the fungus. To identify genes in A. roxburghii that were differentially expressed during the symbiosis with AR-18, we used the differential display reverse transcription polymerase chain reac- tion (DDRT-PCR) method to compare the transcriptomes between seedlings inoculated with the fungus and control seedlings. We amplified 52 DDRT-PCR bands using 15 primer combinations of three anchor primers and five arbitrary primers, and nine bands were re-amplified by double primers. Reverse Northern blot analyses were used to further screen the bands. Five clones were up-regulated in the symbiotic interaction, including genes encoding a uracil phosphoribosyltransferase (UPRTs; EC 2.4.2.9) and a hypothetical protein. One gene encoding an amino acid transmembrane transporter was down-regulated, and one gene encoding a tRNA-Lys (trnK) and a maturase K (matK) pseudogene were expressed only in the inoculated seedlings. The possible roles of the above genes, especially the UPRTs and marK genes, are discussed in relation to the fungal interaction. This study is the first of its type in A. roxburghii. 展开更多
关键词 Anoectochilus roxburghii orchid mycorrhizal symbiosis Epulorhiza sp. differential display-PCR (DD-PCR) gene screening
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