Engineering for Drought Tolerance in Horticultural and Ornamental Plants

Abstract

Drought is one of the major factors limiting plant growth and productivity. Plant adaptation to drought is dependent on molecular networks for drought perception, signal transduction, expression of a subset of genes and production of metabolites that protect and maintain the structure of cellular components. In general, the drought response pathways can be classified into two categories: one is dependent on the stress hormone abscisic acid (ABA) and the other is ABA-independent. Many genes in these pathways have been identified, thereby providing guidance in choosing genes for engineering of drought tolerance. The review highlights the genes that mediate drought response and tolerance, and discusses lessons learned from engineering for drought tolerance in model plants, such as Arabidopsis, rice and tobacco. Because success of drought tolerance engineering is dependent on not only protein coding regions but also appropriate promoters, this article also reviews the promoters that are crucial for successful engineering of stress tolerance.     

Drought Tolerance in Rice: Focus on Recent Mechanisms and Approaches

Drought stress is a serious limiting factor to rice production,which results in huge economic losses.It is becoming a more serious issue with respect to the global climate change.Keeping in view of the current and forecasted global food demand,it has become essential to enhance the crop productivity on the drought-prone rainfed lands with priority.In order to achieve the production target from rainfed areas,there is a requirement of rice varieties with drought tolerance,and genetic improvement for drought tolerant should be a high priority theme of research in the future.Breeding for drought tolerant rice varieties is a thought-provoking task because of the complex nature and multigenic control of drought tolerant traits would be a major bottleneck for the current research.A great progress has been made during last two decades in our understanding of the mechanisms involved in adaptation and tolerance to drought stress in rice.In this review,we highlighted the recent progresses in physiological,biochemical and molecular adaptation of rice to drought tolerance.A brief discussion on the molecular genetics and breeding approaches for drought tolerance in rice will be focused for the future crop improvement program for development of drought tolerant rice varieties.     

RNA干涉下调RACK1基因表达增强水稻抗旱能力

 RACK1是一种多功能支架蛋白,广泛参与植物生长发育过程的调节。利用RNA干涉技术抑制水稻RACK1基因的表达,分析了RACK1基因在响应干旱胁迫中的功能。实时定量PCR对获得的转基因植株的RACK1基因表达分析结果表明,转基因水稻RACK1基因表达受抑制程度达50%左右。与非转基因水稻（对照）相比,转基因水稻耐干旱能力显著强于对照,其膜的过氧化酶和丙二醛的产生显著低于对照,而超氧化物歧化酶活性极显著高于对照。表明RACK1蛋白质调节水稻对干旱胁迫的耐性,并且这种调节在很大程度上与植株体内的氧化还原系统有关。     

栽培稻抗旱性研究的现状与策略

 水资源短缺正成为制约我国农业发展的重要因素。培育抗旱的栽培稻品种并实现水稻旱作，不但可在很大程度上节约水资源，而且有利于增产稳产，节约能源和减少环境污染。抗旱性包括逃旱性、避旱性、耐旱性和复原抗旱性。形态生理学的研究揭示出大量的与栽培稻抗旱性有关的形态特征和生理特性，如根系和叶片性状、生育期、渗透调节、脱落酸含量与栽培稻抗旱性密切相关，且已利用分子标记对上述性状进行了基因定位（QTL）研究。旱稻品种改良也已取得重大进展。在进行抗旱品种改良的基础上，通过引进相应的栽培技术，节水种植，实现水稻旱作，并达到稳产与增产的目的，是抗旱性研究的战略目标。在增产、稳产和优质的前提下，以培育耐旱性极强的水稻（或旱稻）为中心，建立有代表性的抗旱性研究基地, 进一步加强稻属抗旱基因资源的发掘和创新、抗旱生理学和遗传学的研究、利用现代生物技术实现不同物种间抗旱基因的转移、建立节水种植栽培技术新体系是目前抗旱性研究的主要内容。     

Down-Regulated Expression of RACK1 Gene by RNA Interference Enhances Drought Tolerance in Rice

The receptor for activated C-kinase 1 (RACK1) is a highly conserved scaffold protein with versatile functions, and plays important roles in the regulation of plant growth and development. Transgenic rice plants, in which the expression of RACK1 gene was inhibited by RNA interference (RNAi), were studied to elucidate the possible functions of RACK1 in responses to drought stress in rice. Real-time PCR analysis showed that the expression of RACK1 in transgenic rice plants was inhibited by more than 50%. The tolerance to drought stress of the transgenic rice plants was higher as compared with the non-transgenic rice plants. The peroxidation of membrane and the production of malondialdehyde were significantly lower, and the superoxide dismutase activity in transgenic rice plants was significantly higher than those in non-trangenic rice plants. It is suggested that RACK1 negatively regulated the redox system-related tolerance to drought stress of rice plants.     

Gene expression microarrays and their application in drought stress research

Initial physiological adjustments in response to drought stress lead to drastic changes in gene expression. The traditional approaches of assessing such drought-induced changes in gene expression involve measuring the differences in mRNA levels of one or few genes at a time. DNA expression microarray technology is a powerful tool that can monitor changes in expression of a large number of genes simultaneously. Expression microarrays also provide new insights into physiological and biochemical pathways of drought tolerance, and thus can lead to identification of novel candidate genes that can rapidly advance breeding for drought tolerance. This review describes the basic principles and potential applications of gene expression microarrays in understanding and improving drought tolerance in plants. A case study is presented involving hybridization of field-grown panicle samples from drought tolerant and susceptible rice germplasm targets with probes from a normalized panicle cDNA library. Results indicate contrasting drought responses among both upland versus lowland-adapted cultivars and also between traditional and improved upland types.     

水稻耐盐基因定位与克隆及品种耐盐性分子标记辅助选择改良研究进展

土壤盐渍化严重制约水稻生产发展,提高耐盐性已成为水稻育种的重要目标之一。挖掘水稻耐盐新基因,解析其分子作用机制可以为水稻耐盐性遗传改良奠定基础。本文从定位群体、耐盐性鉴定时期和鉴定方法、耐盐性评价指标、鉴定到的耐盐QTL、耐盐QTL的精细定位和图位克隆等方面,总结了近年来水稻耐盐QTL定位研究中所取得的进展;介绍了水稻耐盐/盐敏感突变体筛选和基因克隆以及耐盐性关联分析的研究近况;并对水稻耐盐性分子标记辅助选择改良的现状作了概述。     

A β-ketoacyl-CoA Synthase OsCUT1 Confers Increased Drought Tolerance in Rice

Drought stress is one of the major environmental factors affecting crop growth and productivity. Cuticular wax plays essential roles in protecting plants from environmental stress via forming a hydrophobic barrier on leaf epidermis. In this study, we analyzed nine members (OsCUT1?OsCUT9) of β-ketoacyl-CoA synthase, the rate-limiting key enzyme for cuticular wax synthesis in rice by homology search and domain prediction. The expression levels of OsCUT genes under different abiotic stresses were investigated and OsCUT1 down-regulated by abiotic stress was selected for further function validation. Compared to the wild type, overexpression of OsCUT1 (OX-OsCUT1) exhibited significantly increased drought resistance. Epicuticular wax was increased on the leaf surface of OX-OsCUT1 and the chlorophyll leaching experiment showed that the cuticular permeability was decreased in the OX-OsCUT1 plants. Moreover, overexpression of OsCUT1 didn’t result in the significant changes of major agronomic traits. In total, these results suggested that OsCUT1 is a promising gene for engineering rice plants with enhanced drought tolerance.     

Identification and Characterization of Genes Responsible for Drought Tolerance in Rice Mediated by Pseudomonas fluorescens

Drought is one of the major abiotic stresses which adversely affect crop plants limiting growth and yield potential.Structural and functional characterization of drought stress-induced genes has contributed to a better understanding of how plants respond and adapt to the drought stress.In the present study,differential display technique was employed to study the gene expression of rice plants at the reproductive stage that were subjected to drought stress by withholding water,Pseudomonas fluorescens strain(Pf1) treated plants subjected for drought stress by withholding water and control(well-watered).Differentially expressed c DNAs of six genes(COX1,PKDP,b ZIP1,AP2-EREBP,Hsp20 and COC1) were identified,cloned and sequenced.Real-time q PCR analysis showed that all the six genes were upregulated in drought-stressed plants treated with Pf1.This revealed that the remarkable influence of Pf1 colonization leads to drought tolerance at the reproductive stage.These results showed that high levels of gene expression in plants lacking adequate water can be remarkably influenced by Pf1 colonization,which might be a key element for induced systemic tolerance by microbes.     

水稻类钙调磷酸酶B亚基蛋白基因OsCBL8功能的初步研究

 以转正义或反义OsCBL8基因的T3代株系及非转基因对照为材料, 研究了转OsCBL8基因对水稻农艺性状、内源OsCBL8基因表达和幼苗耐盐、耐旱及耐冷性的影响。结果表明, 供试的16个转基因株系的株高、单株分蘖数、单株有效穗数、穗长和百粒重等性状与对照无显著差异, 每穗实粒数和结实率则大都显著或极显著低于对照。部分转基因株系的半定量RT PCR结果表明, 转正义基因显著增强了植株中OsCBL8基因的表达, 而转反义基因则对内源OsCBL8基因的表达具有一定的抑制作用。在盐、干旱和低温等非生物胁迫条件下的生长实验发现, 1个转正义基因株系的耐盐性得到显著增强, 1个转反义基因株系的耐旱性得到显著增强。 讨论了OsCBL8基因在水稻耐非生物逆境方面可能的机理。     

CbDREB1A基因表达载体构建及转化水稻光温敏核不育系的研究

为了增强荠菜DREB1A基因在转基因水稻中的表达,构建了由Ubi启动子驱动的植物表达载体pUΩCbDREB3300.通过基因枪转化法将CbDREB1A基因导入水稻光温敏核不育系4008S,获得5个抗干旱胁迫的再生植株.运用PCR及Southern杂交技术对抗性再生植株进行鉴定,结果显示CbDREB1A基因已整合到水稻基因组中.干旱胁迫后转基因水稻植株脯氨酸含量显著高于对照,显示耐旱性增强.     

东乡野生稻重要耐逆性状及其分子机制研究进展

东乡野生稻是全球分布最北的普通野生稻,具有丰富的抗逆性状,其育种利用价值高。本文综述了近年来东乡野生稻在耐冷、耐旱及抗虫等重要耐逆性状及其分子机制研究方面取得的进展,以期为水稻耐逆性的进一步研究提供依据。     

一个水稻NADPH氧化还原酶相似基因的克隆及表达分析

采用荧光差显(fluorescent differential display,FDD)技术,比较分析了干旱处理与未处理水稻叶片及根中的mRNA表达,并利用H. A. Yellow PAGE(含0.1% H. A. Yellow)再分离与大矩阵(macroarray)筛选相结合的方法,发现1个与拟南芥NADPH氧化还原酶高度相似(96%)的差异片段。该基因的cDNA全长为1423 bp,编码一个具有345个氨基酸残基的多肽。在正常情况下该基因表达量很低,而在干旱处理中高表达。讨论了干旱胁迫下NADPH氧化还原酶基因的可能功能。     

Cloning and Expression Analysis of OsNADPH1 Gene from Rice in Drought Stress Response

Drought resistance in rice has long been one of the important scientific research programs in cereal crops [1-3]. At present, the finding of novel drought genes is common in rice genetic resource development and varietal improvement. According to the action modes, the drought resistance genes can be divided into two classes [4]. The first class constitutes functional genes coding for such enzymes as late embryogenesis abundant protein (LEA) [5], △1-pyrroline-5-carboxylate synthetase (P5CS) …     

OsABT,a Rice WD40 Domain-Containing Protein,Is Involved in Abiotic Stress Tolerance

Plant growth and crop productivity are severely affected by abiotic stress on a global scale. WD40 repeat-containing proteins play a significant role in the development and environmental adaptation of eukaryotes. In this study, OsABT, a stress response gene, was cloned from rice (Oryza sativa L. cv. Nipponbare). OsABT encodes a protein containing seven WD40 domains. Expression analysis revealed that the OsABT gene was first up-regulated and then down-regulated following treatment with abscisic acid (ABA) and NaCl, but was down-regulated when treated with PEG8000. Subcellular localization results showed that OsABT was located in the cytoplasm and nucleus of Arabidopsis roots. OsABT transgenic Arabidopsis showed significantly increased tolerance to ABA and salt stress during plant seedling development. However, the transgenic lines were more sensitive to drought stress. Moreover, OsABT can interact with OsABI2, a component of ABA signaling pathway. These results showed that OsABT plays a positive regulatory role in response to salt stress and a negative role in response to drought stress in Arabidopsis.     

实时PCR定量分析干旱胁迫下水稻糖原合成酶激酶基因表达差异

通过实时荧光定量PCR进行相对定量分析,得到水稻IR64糖原合成酶激酶（glycogen synthase kinase, GSK）基因在不同生育期、不同组织干旱胁迫处理前后的表达差异。 叶片中GSK基因为组成型表达,各生育期干旱胁迫时诱导合成增加,尤其从分蘖期到抽穗期表达更为显著,说明这段生长期叶片对水分缺乏最敏感;苗期根系处理后GSK表达明显下降,叶片处理前后GSK表达无明显变化。由此可见,GSK基因的表达在不同的发育时期、不同的组织中呈现差异。