水稻耐盐/碱性鉴定评价方法

土壤盐/碱化是盐/碱稻作区水稻生产稳定发展的主要限制因素.为了提高水稻耐盐/碱性,扩大水稻种植面积,减轻盐/碱胁迫导致的水稻减产,许多学者广泛开展了水稻耐盐/碱性的基因型差异、生理生化、遗传及定位等研究,并取得了显著成绩.但国内在耐盐/碱性鉴定评价方法方面还缺乏统一标准,这影响着水稻耐盐/碱性研究的深入开展.本文阐述了国内外至今所采用的水稻耐盐/碱性鉴定方法、耐盐/碱指标和分级标准等,以期为我国水稻耐盐/碱性鉴定评价技术规范的制定以及水稻耐盐/碱性种质资源鉴定、生理生化分析和遗传育种提供参考依据.     

转基因技术在作物抗旱改良中的应用

浙江大学原子核农业科学研究所王忠华、谢建坤、夏英武等研究人员对此课题作了研究。他们以不同靶标基因为例 ,分不同作用机制 (渗透调节和清除氧自由基等 ) ,介绍了近 1 0年来国内外利用转基因技术改良作物耐盐和抗旱性的研究进展 ,为我国北方农业作物抗旱和耐盐性研究与应用提供了参考资料。他们利用转基因的模式作物烟草到直接用粮食作物水稻的抗旱和耐盐基因水稻在生产中应用虽然还需要进一步深入研究 ,但目前国内外农业和植物科学工作者已确定作物的渗透调节、清除活性氧和LEAP(Laeembryogensis abu ndantproteins)蛋白等 3大抗旱与…     

He-Ne激光对盐胁迫下水稻幼苗抗氧化活性的影响

为减轻盐胁迫对植物造成的伤害,本研究采用He-Ne激光对盐胁迫下的水稻进行辐照处理,选用水稻9311作为对照、耐盐海稻86作为研究对象,以0.5%的氯化钠进行盐胁迫,He-Ne激光(辐照计量5 m W·mm-2,波长632.8 nm)进行照射。设置了对照组(CK),盐胁迫组、盐胁迫和激光复合处理组(分别为L、BL组),进行了水稻抗氧化活性方面的研究。结果表明:与对照组(CK)比较,He-Ne激光对水稻的盐胁迫有一定的缓解作用,其中对水稻9311的作用最明显,表现为激光处理提高了水稻幼苗中过氧化物酶(POD)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)的活性和减缓了丙二醛(MDA)含量的增速;同时也说明新选育的海稻86耐盐性高于水稻9311。因此,一定剂量的He-Ne激光可以提高了水稻的耐盐性,这为抗盐胁迫及耐盐植物的育种提供了途径和理论依据。     

盐藻蛋白质组的研究进展

盐藻是一种公认的能耐受高盐度的海藻,是研究植物高盐适应的模式生物.高通量的蛋白质组学为人们深入探讨盐藻的耐盐机制提供了强有力的工具.就蛋白质组及其主要技术、盐藻蛋白质组的研究概况和研究展望作一简要综述.     

植物响应盐胁迫组学研究进展

盐胁迫对植物生长的影响主要表现在离子毒害、渗透胁迫以及次级氧化胁迫等,植物遭受盐胁迫时迅速启动相关基因,进行转录调控,进而合成相应蛋白质来控制代谢物合成和离子转运以调节渗透平衡。随着现代分子生物学迅速发展,对植物耐盐机理研究也深入到了转录组、蛋白质组、代谢组及离子组等水平,"组学"研究为耐盐基因鉴定及标志性代谢物的挖掘等提供了有力手段。该文对近年来国内外有关转录组学、蛋白质组学、代谢组学、离子组学的主要研究方法及在盐胁迫中的应用研究进展进行综述,以揭示植物耐盐机理,为优良耐盐碱植物的筛选与培育提供支持。     

水稻分蘖期耐盐性鉴定评价方法确立及种质筛选

分蘖期是水稻由秧田生长向本田生长过渡的营养生长关键时期。在盐渍化土壤上种植水稻,分蘖期耐盐性对水稻能否正常生长至关重要。合适的耐盐性鉴定方法是水稻耐盐种质筛选的前提。本研究目的在于通过水稻分蘖期盐浓度梯度筛选试验,明确水稻分蘖期耐盐性鉴定最适盐胁迫浓度和调查时间;利用确立的鉴定方法,筛选出水稻分蘖期耐盐种质。本研究选取具有不同程度耐盐性的26份水稻品种,于2015年和2016年分别在10个和7个盐浓度梯度下,以盐害等级为分蘖期耐盐评价指标,开展水稻分蘖期耐盐性鉴定评价试验。结果表明,在0.5%盐浓度下,处理4～6周后供试种质的盐害等级差异表现为最大,因此认为0.5%盐浓度是水稻分蘖期耐盐性鉴定的最适盐胁迫浓度,盐处理后4～6周是水稻分蘖期盐害等级调查的最佳调查时间。参照国际水稻所提出的水稻分蘖期耐盐性评价标准,本研究对其进行了改进,提出以单株为单位进行水稻分蘖期耐盐性评价方法,利用平均盐害等级评估分蘖期耐盐性。该方法能够方便、准确地评价水稻耐盐性强弱。依据上述确立的水稻分蘖期耐盐性鉴定评价方法,本研究于2017-2019年期间对2886份来自不同生态区域的水稻种质进行分蘖期耐盐性鉴定评价,鉴定筛选出137份耐盐种质,为水稻耐盐遗传机制和育种研究提供资源支撑。     

耐盐水稻种质资源的筛选

用0.8% NaCl溶液和国际水稻所水稻耐盐性9级评价方法对前人选留的38份水稻耐盐种质资源(用0.5%NaCl盐土筛选)进行了重复筛选,以期获得极端耐盐水稻种质资源.同时研究了不同盐浓度对水稻发芽率的影响.结果表明:高浓度盐溶液显著抑制水稻种子的发芽率,但品种间有较大差异;筛选出苗期极端耐盐品种6份,其中1级1份,2级4份,3级1份;极端耐盐水稻品种苗高的增长受高浓度盐溶液抑制,但品种间差异不显著;而盐溶液对极端耐盐水稻品种出叶速率的影响因品种而异.     

盐胁迫对4个水稻种质抗逆性生理的影响

为筛选水稻(Oryza sativa)耐盐种质资源,研究了水稻耐盐相关生理指标。结果表明,盐胁迫下,耐盐种质Pokkali、Fl478、JX99的电导率较低,细胞膜透性较小,叶片叶绿素含量较高,受盐害程度较弱。盐胁迫促进了丙二醛的大量积累,加剧了细胞膜脂过氧化程度,而耐盐种质的丙二醛积累较少,但高盐胁迫下3个耐盐种质的丙二醛含量升高,破坏了细胞膜的完整性。感盐种质R29的临界饱和亏的增幅较大,耐盐种质相对含水量降幅小。应用模糊隶属函数法综合评价,4个水稻种质的耐盐性依次为JX99PokkaliFL478IR29。     

水稻全生育期耐盐性鉴定评价方法研究北大核心CSCD

土壤盐渍化是危害水稻生产的重要非生物胁迫因素,而遗传改良是提高水稻耐盐性的有效途径之一。全生育期耐盐性是对水稻各生育时期耐盐性的综合反映。科学、准确、高效的水稻全生育期耐盐性鉴定评价方法是水稻耐盐遗传改良必要条件,也是正确判别水稻种质耐盐真实性的关键所在。本研究挑选19份不同耐盐性水稻种质作为研究材料,在正常环境和0.2%、0.3%、0.4%、0.5%、0.6%、0.7%盐处理浓度下,调查11个农艺性状并计算其耐盐系数,运用品种间四分位差法分析,明确水稻全生育期耐盐性鉴定最适浓度。品种间四分位差最大的盐浓度被认为是盐处理最佳浓度;综合多种多元统计分析方法,包括主成分分析、隶属函数分析、回归分析、聚类分析,对水稻全生育期耐盐性鉴定合理评价方法进行探索。结果表明,0.3%盐胁迫浓度下多数性状耐盐系数品种间四分位差最大,0.3%盐胁迫浓度是水稻全生育期耐盐性鉴定最适浓度。主成分分析结果表明,11个农艺性状的耐盐系数可简化为3个主成分。利用主成分贡献率和隶属函数分析,可进一步将3个主成分值简化为水稻耐盐性综合评价指标D值。D值能够简便、准确的评价水稻种质的耐盐性。本研究还利用逐步回归分析,建立D值与11个农艺性状耐盐系数最优线性回归方程:D=-0.365+0.647PL+0.152GP+0.274TW。从该方程可知穗长、穗粒数和总干物重耐盐系数是影响D值的关键指标。利用回归分析建立的模型,可准确完成对D值预测。本研究利用聚类分析,将19份水稻种质耐盐性划分为5个等级,对应于水稻5个耐盐等级划分,即极强、强、中、弱、极弱,可作为其他水稻种质耐盐性评价重要参考。     

水稻全生育期耐盐性鉴定评价方法研究

土壤盐渍化是危害水稻生产的重要非生物胁迫因素,而遗传改良是提高水稻耐盐性的有效途径之一。全生育期耐盐性是对水稻各生育时期耐盐性的综合反映。科学、准确、高效的水稻全生育期耐盐性鉴定评价方法是水稻耐盐遗传改良必要条件,也是正确判别水稻种质耐盐真实性的关键所在。本研究挑选19份不同耐盐性水稻种质作为研究材料,在正常环境和0.2%、0.3%、0.4%、0.5%、0.6%、0.7%盐处理浓度下,调查11个农艺性状并计算其耐盐系数,运用品种间四分位差法分析,明确水稻全生育期耐盐性鉴定最适浓度。品种间四分位差最大的盐浓度被认为是盐处理最佳浓度;综合多种多元统计分析方法,包括主成分分析、隶属函数分析、回归分析、聚类分析,对水稻全生育期耐盐性鉴定合理评价方法进行探索。结果表明,0.3%盐胁迫浓度下多数性状耐盐系数品种间四分位差最大,0.3%盐胁迫浓度是水稻全生育期耐盐性鉴定最适浓度。主成分分析结果表明,11个农艺性状的耐盐系数可简化为3个主成分。利用主成分贡献率和隶属函数分析,可进一步将3个主成分值简化为水稻耐盐性综合评价指标D值。D值能够简便、准确的评价水稻种质的耐盐性。本研究还利用逐步回归分析,建立D值与11个农艺性状耐盐系数最优线性回归方程:D=-0.365+0.647PL+0.152GP+0.274TW。从该方程可知穗长、穗粒数和总干物重耐盐系数是影响D值的关键指标。利用回归分析建立的模型,可准确完成对D值预测。本研究利用聚类分析,将19份水稻种质耐盐性划分为5个等级,对应于水稻5个耐盐等级划分,即极强、强、中、弱、极弱,可作为其他水稻种质耐盐性评价重要参考。     

水稻高温热害发生机理与耐高温遗传基础研究

在全球气候变暖的大环境下,极端天气频繁发生,高温已经成为制约水稻产量和品质的主要因素之一。培育耐高温的水稻品种是防止高温热害最简便有效的途径。现代分子技术的发展和水稻功能基因组研究的深入,为水稻耐高温遗传机制的剖析提供了有效手段。本文综述了近年来水稻高温热害的发生机理和耐高温遗传基础方面的研究进展,包括耐高温QTL定位与克隆、转录组研究、蛋白组研究等,以期为深入剖析水稻耐高温的分子机制和培育耐高温的新品种提供有益参考。     

适于蛋白双向电泳的水稻叶片样品提取方法初探

在水稻基因组测序完成后,利用蛋白质组学技术揭示水稻基因功能的研究,已成为水稻分子生物学研究的热点之一。水稻叶片作为DNA研究的便利材料被经常使用,但对蛋白质研究来说,占叶片全蛋白50%～60%的核酮糖二磷酸羧化酶（RuBP羧化酶）对低丰度蛋白常常造成掩盖。以水稻叶片为材料,用不同浓度的聚乙二醇（PEG）去除叶片中RuBP羧化酶。通过SDS-PAGE垂直电泳比较发现,浓度为17%的PEG对去除RuBP羧化酶效果最好,所获得的蛋白质样品可以得到质量较高的双向电泳图谱。     

Update and challenges on proteomics in rice

Rice is not only an important agricultural resource but also a model plant for biological research. Our previous review highlighted different aspects of the construction of rice proteome database, cataloguing rice proteins of different tissues and organelle, differential proteomics using 2-DE and functional characterization of some of the proteins identified (Komatsu, S., Tanaka, N., Proteomics 2005, 5, 938-949). In this review, the powerfulness and weaknesses of proteomic technologies as a whole and limitations of the currently used techniques in rice proteomics are discussed. The information obtained from these techniques regarding proteins modification, protein-protein interaction and the development of new methods for differential proteomics will aid in deciphering more precisely the functions of known and/or unknown proteins in rice.     

基于Make2D-DB II构建水稻二维电泳-质谱联动数据库

基于Make2D—DBII软件包构建了一个水稻二维电泳-质谱联动数据库Rice2DDB,介绍了该数据库的构建方法和步骤,为快速发展的水稻蛋白质组学研究提供了数据管理和交流的平台,对其他物种的蛋白质组学研究也具有参考价值。     

Use of proteomics to understand seed development in rice

Rice is an important cereal crop and has become a model monocot for research into crop biology. Rice seeds currently feed more than half of the world's population and the demand for rice seeds is rapidly increasing because of the fast‐growing world population. However, the molecular mechanisms underlying rice seed development is incompletely understood. Genetic and molecular studies have developed our understanding of substantial proteins related to rice seed development. Recent advancements in proteomics have revolutionized the research on seed development at the single gene or protein level. Proteomic studies in rice seeds have provided the molecular explanation for cellular and metabolic events as well as environmental stress responses that occur during embryo and endosperm development. They have also led to the new identification of a large number of proteins associated with regulating seed development such as those involved in stress tolerance and RNA metabolism. In the future, proteomics, combined with genetic, cytological, and molecular tools, will help to elucidate the molecular pathways underlying seed development control and help in the development of valuable and potential strategies for improving yield, quality, and stress tolerance in rice and other cereals. Here, we reviewed recent progress in understanding the mechanisms of seed development in rice with the use of proteomics.     

Biotic Stress-Responsive Rice Proteome: An Overview

Biotic stresses affect the plant growth, seed quality, and crop yield. The monocot model rice crop plant is no exception and is affected by a variety of biotic stress factors. High-throughput proteomics approaches are being applied in rice for the past several years to exploit better understanding the biotic stresses-responsive regulatory mechanisms. A large number of proteins responsive to biotic stresses, including pathogens and herbivores, have been cataloged. Cataloged proteins mainly belong to functional categories into metabolism, energy, defense mechanisms, and signaling. To date, majority of these proteins have not been functionally characterized yet, except the pathogen-related 10 protein, PBZ1. This review will briefly summarize and discuss: (1) the proteomics-based investigation of biotic stress-responsive proteins in rice and (2) increasing importance of proteomics approach in defense biology and engineering the next-generation rice/crop plants.     

Proteome profiling highlights mechanisms underlying pigment and tocopherol accumulation in red and black rice seeds

Rice is a major component of the human diet and feeds more than 50 million people across the globe. We previously developed two pigmented rice cultivars, Super-hongmi (red seeds) and Super-jami (black seeds), that are highly rich in antioxidants and exhibit high levels of radical scavenging activities. However, the molecular mechanism underlying the accumulation of pigments and different antioxidants in these rice cultivars remains largely elusive. Here, we report the proteome profiles of mature Super-hongmi and Super-jami seeds, and compared them with the Hopum (white seeds) using a label-free quantitative proteomics approach. This approach led to the identification of 5127 rice seed proteins of which 1628 showed significant changes in the pigmented rice cultivar(s). The list of significantly modulated proteins included a phytoene desaturase (PDS3) which suggested accumulation of ζ-carotene in red seeds while the black seeds seem to accumulate more of anthocyanins because of the higher abundance of dihydroflavonol 4-reductase. Moreover, proteins associated with lignin and tocopherol biosynthesis were highly increased in both red and black cultivars. Taken together, these data report the seed proteome of three different colored rice seeds and identify novel components associated with pigment accumulation in rice.     

水稻功能基因组学研究

水稻是迄今为止第一个被测序的农作物。随着水稻基因组测序计划的完成,以功能基因组学研究为标志的后基因组时代已经到来。综述了水稻功能基因组学的工作进展与方法,主要包括:表达序列标签(EST)c、DNA微阵列和DNA芯片、蛋白质组学、生物信息学和反向遗传学等新方法。     

Rejuvenating rice proteomics: facts, challenges, and visions

Proteomics is progressing at an unprecedented pace, as can be exemplified by the progress in model organisms such as yeast, bacteria, and mammals. Proteomics research in plants, however, has not progressed at the same pace. Unscrambling of the genome sequences of the dicotyledoneous Arabidopsis thaliana (L.) and monocotyledoneous rice (Oryza sativa L.) plant species, respectively, has made them accessible reference organisms to study plant proteomics. Study of these two reference plants is expected to unravel the mystery of plant biology. Rice, a critically important food crop on the earth, has been termed a "cornerstone" and the "Rosetta stone" for functional genomics of cereal crops. Here, we look at the progress in unraveling rice proteomes and present the facts, challenges, and vision. The text is divided into two major parts: the first part presents the facts and the second part discusses the challenges and vision. The facts include the technology and its use in developing proteomes, which have been critically and constructively reviewed. The challenges and vision deal with the establishment of technologies to exhaustively investigate the protein components of a proteome, to generate high-resolution gel-based reference maps, and to give rice proteomics a functional dimension by studying PTMs and isolation of multiprotein complexes. Finally, we direct a vision on rice proteomics. This is our third review in series on rice proteomics, which aims to stimulate an objective discussion among rice researchers and to understand the necessity and impact of unraveling rice proteomes to their full potential.     

植物响应病原菌胁迫的蛋白质组学研究进展

随着拟南芥、水稻等模式植物基因组测序的完成,植物基因组学的研究重点已经转变为功能基因组学研究。蛋白质组学成为后基因组时代的重要研究手段,它有助于从分子水平上了解植物功能。主要介绍了双向电泳技术、生物质谱、蛋白质质谱数据的生物信息学分析等蛋白质组学研究的主要技术手段及植物应答病原菌胁迫的蛋白质组学研究进展,并对蛋白质组学在研究植物抗病机制方面的应用前景做出展望。