Suppression Subtractive Hybridization Analysis of Genes Regulated by Application of Exogenous Abscisic Acid in Pepper Plant (Capsicum annuum L.) Leaves under Chilling Stress.
Suppression Subtractive Hybridization Analysis of Genes Regulated by Application of Exogenous Abscisic Acid in Pepper Plant (Capsicum annuum L.) Leaves under Chilling Stress.
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DOI：
10.1371/journal.pone.0066667
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Li DW
中科院分区：
文献类型：
--
作者：
Guo WL;Chen RG;Gong ZH;Yin YX;Li DW
低温是限制辣椒(Capsicum annuum L.)在非热带地区的冬季和早春生产。外源脱落酸(ABA)能有效缓解辣椒叶片的冷害症状，如枯萎和坏死病的形成，但其分子机制尚不清楚。本研究的目的是利用抑制消减杂交(SSH)的方法，在6℃培养48h的辣椒幼苗中，找出ABA处理后差异表达上调或下调的基因。共分离、聚类、组装了235个优质EST，共73个非基因片段，包括18个重叠群和55个单态。共有37个(50.68%)的单基因与非冗余数据库中已知功能的基因具有相似性，另外36个(49.32%)的单基因与非冗余数据库中的已知功能的基因具有较低的相似性或未知功能。基因本体论分析表明，37个UNIGNES可分为9个功能类别。用定量RT-PCR分析了18个基因的表达谱，其中10个基因在ABA处理的幼苗中的表达水平至少是水分处理(对照)幼苗在低温胁迫下的2倍。相比之下，在ABA处理的幼苗中，其他8个基因在低温胁迫下表达下调，表达水平仅为对照幼苗的三分之一或更少。这些结果表明，ABA对低温胁迫下辣椒植株的基因有正向和负向调节作用。
Low temperature is one of the major factors limiting pepper (Capsicum annuum L.) production during winter and early spring in non-tropical regions. Application of exogenous abscisic acid (ABA) effectively alleviates the symptoms of chilling injury, such as wilting and formation of necrotic lesions on pepper leaves; however, the underlying molecular mechanism is not understood. The aim of this study was to identify genes that are differentially up- or downregulated in ABA-pretreated hot pepper seedlings incubated at 6°C for 48 h, using a suppression subtractive hybridization (SSH) method. A total of 235 high-quality ESTs were isolated, clustered and assembled into a collection of 73 unigenes including 18 contigs and 55 singletons. A total of 37 unigenes (50.68%) showed similarities to genes with known functions in the non-redundant database; the other 36 unigenes (49.32%) showed low similarities or unknown functions. Gene ontology analysis revealed that the 37 unigenes could be classified into nine functional categories. The expression profiles of 18 selected genes were analyzed using quantitative RT-PCR; the expression levels of 10 of these genes were at least two-fold higher in the ABA-pretreated seedlings under chilling stress than water-pretreated (control) plants under chilling stress. In contrast, the other eight genes were downregulated in ABA-pretreated seedlings under chilling stress, with expression levels that were one-third or less of the levels observed in control seedlings under chilling stress. These results suggest that ABA can positively and negatively regulate genes in pepper plants under chilling stress.
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