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鉴定与黄瓜高磷积累相关的上调基因。

Characterization of upregulated genes associated with high phosphorus accumulation in cucumber.

机构信息

Department of Biology, Western Kentucky University, Bowling Green, KY 42101, USA.

出版信息

Physiol Plant. 2011 Dec;143(4):344-54. doi: 10.1111/j.1399-3054.2011.01512.x. Epub 2011 Oct 7.

Abstract

Excessive application of phosphorus (P)-rich manures to agricultural lands often results in P-accumulation in soils leading to water pollution through runoffs and leaching. Use of suitable plant species that can extract and sequester excess P from soil into their biomass is an effective method of remediation of P-contaminated soils. Knowledge on the molecular responses of plants to high P-accumulation and tolerance is lacking. Therefore, a suppression subtractive hybridization (SSH) strategy was employed to identify and elucidate the pattern of gene expression related to P-tolerance and accumulation in cucumber (Cucumis sativus L.), a P-accumulator plant. RNA isolated from cucumber grown in high P was used for 'tester' cDNA synthesis and SSH library preparation. A total of 63 cDNAs were identified as showing upregulated expression in this plant in response to high P. No putative function could be assigned to 7 (11%) of the 63 upregulated high P-modulated genes and 11 expressed sequence tags (ESTs) (17%) did not match database entries. The remaining 45 ESTs were grouped into five functional classes. The majority of these ESTs belonged to three groups: 'metabolism', 'protein synthesis/degradation and signaling' and 'cell structure/cell wall'. Only six 'stress/defense'-related ESTs were identified from this library. The results of reverse northern blot analysis was further confirmed and validated through semi-quantitative RT-PCR carried out with representative ESTs identified in this study. The research reported here may contribute to a preliminary understanding of the high P-related gene expression in this P-accumulating plant.

摘要

过量施用富含磷 (P) 的肥料到农田中常常导致土壤中磷的积累,从而通过径流和淋溶导致水污染。利用能够从土壤中提取并将过量磷固定到生物量中的合适植物物种是修复受磷污染土壤的有效方法。然而,人们对植物对高磷积累和耐受的分子反应知之甚少。因此,采用抑制性消减杂交 (SSH) 策略来鉴定和阐明与黄瓜 (Cucumis sativus L.) 中 P 耐受和积累相关的基因表达模式,黄瓜是一种 P 积累植物。从在高 P 下生长的黄瓜中分离 RNA 用于 '测试者' cDNA 合成和 SSH 文库制备。共有 63 个 cDNA 被鉴定为在这种植物中对高 P 表现出上调表达。这 63 个上调的高 P 调节基因中,有 7 个 (11%) 没有功能,11 个表达序列标签 (ESTs) (17%) 与数据库条目不匹配。剩余的 45 个 EST 被分为五个功能类别。这些 EST 中的大多数属于三个组:'代谢'、'蛋白质合成/降解和信号转导'和'细胞结构/细胞壁'。仅从这个文库中鉴定出 6 个与'应激/防御'相关的 EST。通过用本研究中鉴定的代表性 EST 进行半定量 RT-PCR 进行的反向 Northern 印迹分析进一步证实和验证了该结果。本研究的结果可能有助于初步了解这种 P 积累植物中与高 P 相关的基因表达。

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