Profiling of differentially expressed genes in roots of Robinia pseudoacacia during nodule development using suppressive subtractive hybridization.
Profiling of differentially expressed genes in roots of Robinia pseudoacacia during nodule development using suppressive subtractive hybridization.
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利用抑制性消减杂交技术分析刺槐根在根瘤发育过程中差异表达的基因
DOI：
10.1371/journal.pone.0063930
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Wei G
中科院分区：
文献类型：
--
作者：
Chen H;Chou M;Wang X;Liu S;Zhang F;Wei G
背景豆科植物-根瘤菌共生是一个复杂的过程，在宿主植物细胞中通过基因表达网络进行调节。已在豆科草本植物中鉴定出许多在结瘤不同阶段上调的结瘤蛋白基因。然而，尚未报道木本豆科植物如刺槐（Robiniapseudoacacia）中的结节素基因。方法/主要发现 为了鉴定参与假金合欢-紫穗槐中生根瘤菌 CCNWGS0123 共生的结瘤素基因，采用抑制性消减杂交方法来揭示差异表达基因的概况，并构建了两个消减 cDNA 文库，每个文库包含 600 个克隆。然后，通过差异筛选从正向SSH文库中鉴定出114个unigene，并将这些翻译产物的假定功能分为13类。由于对调节基因特别感兴趣，根据逆转录聚合酶链式反应 (RT-PCR) 分析的结果，选择了编码潜在调节蛋白的 21 个上调基因。它们包括九个假定的转录基因、八个假定的翻译后调节基因和四个膜蛋白基因。通过定量RT-PCR进一步分析了这些基因在根瘤发育不同阶段的表达模式。结论 这里提供的数据首次深入了解了 R.pseudoacacia–M 的分子基础。紫穗槐共生。本研究中筛选的许多调控基因揭示了高度的调控复杂性（转录、转录后、翻译和翻译后），这可能对于发展共生至关重要。此外，还讨论了这些基因在刺槐结瘤中的可能作用。
Background Legume-rhizobium symbiosis is a complex process that is regulated in the host plant cell through gene expression network. Many nodulin genes that are upregulated during different stages of nodulation have been identified in leguminous herbs. However, no nodulin genes in woody legume trees, such as black locust (Robinia pseudoacacia), have yet been reported. Methodology/Principal findings To identify the nodulin genes involved in R. pseudoacacia-Mesorhizobium amorphae CCNWGS0123 symbiosis, a suppressive subtractive hybridization approach was applied to reveal profiling of differentially expressed genes and two subtracted cDNA libraries each containing 600 clones were constructed. Then, 114 unigenes were identified from forward SSH library by differential screening and the putative functions of these translational products were classified into 13 categories. With a particular interest in regulatory genes, twenty-one upregulated genes encoding potential regulatory proteins were selected based on the result of reverse transcription-polymerase chain reaction (RT-PCR) analysis. They included nine putative transcription genes, eight putative post-translational regulator genes and four membrane protein genes. The expression patterns of these genes were further analyzed by quantitative RT-PCR at different stages of nodule development. Conclusions The data presented here offer the first insights into the molecular foundation underlying R. pseudoacacia–M. amorphae symbiosis. A number of regulatory genes screened in the present study revealed a high level of regulatory complexity (transcriptional, post-transcriptional, translational and post-translational) that is likely essential to develop symbiosis. In addition, the possible roles of these genes in black locust nodulation are discussed.
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DOI：
10.1073/pnas.1107912108
发表时间：
2011-08-23
影响因子：
11.1
作者：
Capoen, Ward;Sun, Jongho;Oldroyd, Giles E. D.
通讯作者：
Oldroyd, Giles E. D.
影响因子：
12.3
作者：
Andersson A;Keskitalo J;Sjödin A;Bhalerao R;Sterky F;Wissel K;Tandre K;Aspeborg H;Moyle R;Ohmiya Y;Bhalerao R;Brunner A;Gustafsson P;Karlsson J;Lundeberg J;Nilsson O;Sandberg G;Strauss S;Sundberg B;Uhlen M;Jansson S;Nilsson P
通讯作者：
Nilsson P
影响因子：
7.4
作者：
Baima, S;Possenti, M;Morelli, G
通讯作者：
Morelli, G
影响因子：
3.5
作者：
Bloemberg, GV;Wijfjes, AHM;Lugtenberg, BJJ
通讯作者：
Lugtenberg, BJJ
影响因子：
7.2
作者：
Ben-Naim, O;Eshed, R;Lifschitz, E
通讯作者：
Lifschitz, E