水稻叶片早衰突变体的农艺与生理性状研究进展

叶片早衰严重影响水稻产量与品质。研究水稻叶片早衰突变体的农艺及生理性状,有助于了解水稻叶片衰老进程和机理。本文简述了叶片衰老特征和衰老机制,重点综述了水稻叶片早衰突变体的类型、农艺与生理性状研究进展及延缓水稻叶片衰老的栽培调控途径。可为水稻抗衰老品种选育及栽培调控提供理论依据。     

水稻剑叶衰老过程中内源IAA的轭合态变化及其氮素调控效应

目的 揭示水稻抽穗开花后剑叶衰老过程的内源自由态IAA和轭合态IAA含量的动态变化及其基因型间的差异,明确氮素营养供应对叶片衰老过程的延缓效应及其与两种化学态IAA含量之间的关系。方法 以剑叶早衰水稻突变体(psf)与其野生型(浙恢7954)为材料,比较了其在大田条件下剑叶衰老过程中的内源IAA含量、IAA类型和YUCCAs家族基因转录表达量差异,并利用水培种植试验设低氮(LN, 1.45 mmol/L)、正常氮(NN, 2.9 mmol/L)和高氮(HN, 5.8 mmol/L)3个氮浓度水平处理,对不同氮素水平下水稻叶片衰老相关指标与两类IAA含量间的关系进行了分析。结果 早衰突变体psf剑叶中的自由态IAA含量显著低于相同取样时期的野生对照,但前者剑叶中的轭合态IAA含量显著高于后者,且随叶片衰老进程呈较明显的上升趋势;在7个YUCCAs同工型基因中,YUCCA1、YUCCA2、YUCCA3、YUCCA4和YUCCA6)的转录表达量随叶片衰老而下降,但YUCCA7则随叶片衰老呈现出先上升后下降的变化趋势。相同灌浆时期相比,绝大多数YUCCAs在早衰突变体psf剑叶中的转录表达量要低于其野生型对照,说明IAA合成代谢较弱是psf剑叶叶绿素含量迅速下降和早衰症状加剧的重要原因之一。其中,YUCCA7转录表达量随叶片衰老而上升,可能对psf已衰老叶片中的IAA合成起重要调控作用;氮浓度对水稻叶片中的IAA含量及其自由态与共轭态的影响表现为,在叶片开始衰老的初期和前中期,低氮处理(LN)引起剑叶中的自由态IAA含量下降和轭合态IAA含量上升,而高氮处理(HN)可诱导叶片中的自由态IAA含量增加,并导致轭合态IAA含量明显降低,但对已严重衰老的叶片而言(psf在抽穗开花后28 d的剑叶),高氮处理下(HN)的自由态IAA和轭合态IAA含量均低于正常氮处理(NN)和低氮处理(LN)。结论 水稻叶片的衰老进程与其叶片器官中的IAA含量及其化学态之间存在较密切联系。其中,轭合态IAA含量在叶片衰老过程的大幅度提升,是早衰突变体psf 剑叶衰老进程加快的一个重要生理特征,而轭合态IAA含量的下降,可能是高氮处理对水稻叶片衰老进程起到延缓作用的一个重要生理调节因素。     

烟酰胺腺嘌呤二核苷酸(NAD)合成途径和水稻叶片早衰

叶片是植物进行光合作用的主要场所,其衰老由内源遗传发育信号和外界环境胁迫所启动,是一个非常复杂有序的调控过程。烟酰胺腺嘌呤二核苷酸(nicotinamide adenine dinucleotide,NAD)是脱氢酶的辅酶,在糖酵解、糖异生、三羧酸循环以及呼吸链等代谢中发挥着不可替代的作用。最新研究表明,水稻NAD生物合成参与调控沉默信息调控因子Sirtuins的生物活性、组蛋白H3K9去乙酰化、植物激素茉莉酸(JA)和叶片衰老。本文综述了有关水稻叶片衰老的细胞生理特征、Sirtuins酶活、NAD生物合成以及水稻早衰的OsSRT1-NAD调控途径和OsSRT1-Me OH-JA调控途径,以期阐明水稻叶片衰老的分子机理及其调控途径,为高产育种提供相应的理论参考。     

水稻叶片衰老研究进展

叶片的衰老是植物生长发育的最后阶段,是一种基因编码的程序性事件。但在农业生产上,水稻叶片的过早衰老已经成为制约水稻发展的重要因素,研究水稻叶片的早衰对水稻育种改良具有重要意义。本文综述了近年来国内外有关水稻叶片衰老研究结果,主要包括水稻叶片衰老的机理、特征、影响叶片衰老的因子、常用的水稻叶片衰老的生理生化指标等,同时介绍了近年来有关水稻叶片衰老遗传研究进展。     

水稻黄叶早衰突变体osyes1的生理特性和基因定位

【目的】叶片是水稻进行光合作用最重要的器官。叶色突变体是研究光合作用、叶绿素合成代谢和叶绿体发育的重要材料。【方法】利用⁶⁰Co辐射诱变中籼恢复系自选1号,获得黄叶早衰突变体osyes1。幼苗期对突变体和野生型进行外源H₂O₂处理。抽穗期对突变体和野生型叶片进行超氧化物歧化酶活性、过氧化氢酶活性、活性氧含量、丙二醛含量、可溶性蛋白含量、叶绿素含量、净光合速率测定以及透射电镜观察。成熟期考查突变体和野生型的主要农艺性状。以osyes1/02428的F₂群体中的隐性单株为作图群体,利用图位克隆方法定位OsYES1基因。【结果】osyes1的突变性状始于3~4叶期,水稻抽穗后,所有叶片均表现为黄叶衰老症状,致使突变体株高、穗长、每穗粒数及结实率极显著低于野生型对照。与野生型对照相比,突变体幼苗对外源H₂O₂更敏感。生理分析表明,孕穗期野生型倒3叶的叶绿素含量、过氧化物酶和过氧化氢酶活性极显著低于倒2叶和剑叶,而突变体的含量则极显著低于野生型且依次极显著降低;与野生型相比,突变体剑叶、倒2叶和倒3叶的丙二醛、H₂O₂和O₂^-含量极显著增加,而可溶性蛋白含量则相反。遗传分析表明,osyes1受一对隐性核基因控制,利用图位克隆技术将OsYES1基因定位于第7染色体短臂的RM21353与RM21384之间,物理距离为708 kb。【结论】由于osyes1叶片过早黄化衰老导致与产量相关的重要农艺性状显著下降。OsYES1基因定位在第7染色体两个SSR标记(RM21353和RM21384)之间的708 kb的物理区间内。     

水稻新型黄化转绿叶色突变体的遗传分析与育种利用

在育种实践中,叶色变异可作为标记性状,在种子生产和纯度鉴定等方面得到利用。本研究中,对一个水稻新型叶色黄化转绿型自然突变体进行了形态特征观察、遗传分析和育种利用,比较了突变体与野生型的主要农艺性状,利用突变体与正常绿色水稻材料构建遗传群体对该突变体进行经典遗传学分析,通过回交选育了带叶色标记的新不育系。形态观察发现,突变体植株从苗期到分蘖剑叶全展以前的整个叶片生长发育期能持续观察到各个叶片从叶尖向叶基部由黄转绿的发育特性。突变体的主要农艺性状和产量性状与野生型基本一致;遗传分析表明突变体受1对隐性核基因控制。利用突变体与三系不育系回交转育而成的实用型黄化转绿型标记叶色不育系标记性状明显、败育彻底、配合力和可恢复性好。该突变体不同于其它叶色突变体的叶色表达特性具有很好的基础研究和育种利用价值。     

弹性漆改善软木地板漆膜性能的研究

为了探明水稻营养生长阶段的衰老机制,鉴定了一个在抽穗期前多数叶片逐渐死亡的黄色水稻突变体。黄色衰老突变体表型特征为在分蘖期后,其绿色叶片基部随机出现斑点,并逐渐扩散至整个叶片。与之相反,野生型叶片在整个分蘖期均保持绿色。超微结构分析发现,水稻突变体叶片叶绿体类囊体膜和其他细胞器结构在不同衰老时期破裂,严重时导致细胞质皱缩。在叶片衰老过程中,突变体叶片光合活性、叶绿素含量和抗氧化酶活性发生不可逆转的下降。     

水稻叶片衰老基因LPS1的克隆与功能研究

【目的】早衰突变体是研究早衰机制的良好载体,对于探究早衰的遗传机理与作用机制及提高水稻的产量和品质具有重要作用。【方法】本研究利用EMS诱变获得了一个早衰突变体lps1,并对该突变体及其野生型进行表型观察、细胞学及组织化学分析、生理生化分析、遗传分析、基因定位和激素处理。【结果】lps1的叶片从3叶期开始发黄,成熟期株高、有效分蘖数、结实率、千粒重等极显著降低。电镜观察发现lps1叶表面光滑,硅质化突起和叶绿体数目减少、片层结构紊乱。生理生化分析表明lps1中有大量的活性氧积累,同时伴有蛋白质的降解、细胞膜的损伤以及大规模的细胞死亡。遗传分析表明该早衰表型受单隐性核基因控制,并且其在第5染色体上编码了一个泛素结合酶。亚细胞定位结果证明LPS1蛋白在细胞质与核中均有表达。外源激素处理发现,lps1对外源激素的处理更为敏感,且LPS1突变促进了ABA合成相关基因的表达。【结论】LPS1 突变使水稻ABA合成信号途径异常,进而引发H₂O₂等一系列与衰老相关生理指标的异常变动,导致lps1过早衰老,最终造成水稻产量严重降低。     

4PU－30延缓杂交水稻叶片衰老的生理基础

以汕优63连体和离体叶片为材料,研究了4PU-30对水稻叶片衰老进程及叶片内蛋白质、核酸含量和相关酶活力的调控效应。4PU-30能显著地延缓水稻叶片衰老,保绿效果明显。叶片中蛋白质和核酸含量及相关酶活力的变化表明,4PU-30控制叶片衰老过程中蛋白质水解酶和核糖核酸酶比活上升,减缓蛋白质和核酸的降解,是其延缓杂交水稻叶片衰老的主要生理基础之一。     

水稻叶片衰老与超氧化物歧化酶(SOD)

水稻早衰是限制产量和品质提高的最重要因素之一。叶片是水稻最重要的源器官,其功能及衰老对水稻产量与品质有重要影响。叶片衰老特性的研究对提高光合潜力、提高肥料等投入的利用率和经济系数有重要指导意义(Ｗａｄａｙ,1991;Ｔｈｏｍａｓ等,1993)。因此,人们很早以前便已开展水稻叶片衰老生理的研究,特别是在近十几年中发展尤为迅速。但是,有关叶片衰老的机制还远没有搞清楚,特别是对原初伤害机制了解更少。衰老是生物界的普遍现象,是生物发展进化过程中不可分割的一部分。在众多有关水稻叶片衰老的学说中,生物自由基伤害学说引起人们广泛…     

Physiological Characters and Gene Mapping of a Yellow Leaf and Early-senescence Mutant osyes1 in Rice

【Objective】Leaf is a main photosynthetic organ and its color mutants of rice are ideal materials for the study of photosynthesis, chlorophyll biosynthesis and metabolism, and chloroplast development in plants. 【Method】The mutant osyes1 (Oryza sativa yellow-leaf and early senescence 1) was obtained through 60Co γ-radiation treatment of indica cultivar Zixuan 1. The seedlings of osyes1 and its wild type were treated with exogenous H2O2. The SOD and CAT activities, ROS level, MDA content, soluble protein contents, chlorophyll contents, the net photosynthetic rate and chloroplast ultrastructure were analyzed for osyes1 and its wild type leaves at heading stage. The main agronomic traits of osyes1 and its wild type plants were analyzed under field conditions at maturation stage in Hangzhou. The recessive individuals in F2 population derived from osyes1/02428 were used to locate the gene by the map cloning method.【Results】The yellow-leaf and early-senescence symptoms started at 3- or 4-leaf stage, and gradually spread to all of the leaves after heading. Due to the early-senescence of the leaves of the mutant osyes1, its major agronomic traits including plant height, panicle length, grain number per panicle and the seed setting rate were markedly reduced. Moreover, the mutant osyes1 exhibited hyper-sensitivity to exogenous H2O2. The physiological analysis indicated that the contents of chlorophyll and the activities POD and CAT in the third-top leaf was significantly lower than those in the flag leaf and the second-top leaf in wild-type (WT) plants, but all of them in the mutant plants were significantly lower than those in its WT. Compared with the WT plants, the contents of MDA, H2O2 and O2− followed a steady increasing trend in the flag, second-top and third-top leaves of the mutant, while their soluble protein levels were progressively dropping. Genetic analysis confirmed that osyes1 was controlled by a single recessive nuclear gene, which was mapped to a region of 708 kb flanked by two SSR markers (RM21353 and RM21384) on the short arm of chromosome 7. 【Conclusion】In this work, the main agricultural traits were significantly reduced in osyes1 for the yellow leaf and early senescence. OsYES1 was located in a 708 kb range between RM21353 and RM21384 by map-based cloning strategy.     

Characterization of a Novel Gain-of-Function Spotted-Leaf Mutant with Enhanced Disease Resistance in Rice

We here reported the identification and characterization of a novel gain-of-function spotted-leaf 26(Spl26) mutant from an ethylmethylsulfone(EMS)-induced rice cultivar IR64. Spl26 displayed reddishbrown lesions that firstly appeared on the leaf tips at the early tillering stage and spread gradually downward to cover the whole leaf blades that wilted subsequently. The lesion development was lightdependent under natural conditions. Spl26 exhibited impaired photosynthetic capacity with decreased chlorophyll content and lowered photosynthetic parameters which ultimately led to the poor performance of agronomic traits. Severe cell death occurred in Spl26 in accompany with increased malonaldehyde level and membrane ion leakage rate, elevated reactive oxygen species(ROS) accumulation, altered ROS scavenging activities, increased DNA fragmentation and decreased soluble protein levels. Defense responses were activated in Spl26 with enhanced resistance to rice bacterial blight, up-regulation of defense response genes and altered endogenous hormone levels. The spotted-leaf phenotype is controlled by a single dominant nuclear gene localized to a 305 kb region between RM5490 and In Del42 on the short arm of chromosome 7. The data suggested that Spl26 is a novel gain-of-function spotted-leaf mutant with enhanced bacterial disease resistance and immunity-associated premature leaf senescence and would provide the basis for cloning of the target gene.     

控释氮肥调控水稻光合功能和叶片衰老的生理基础

 以杂交水稻组合八两优100和威优46为试验材料，探讨了控释氮肥对水稻光合功能衰退和叶片衰老的调控机制。结果表明，控释氮肥对延缓早、晚稻功能叶的衰老和延长叶片的光合功能期均具有明显的效应。与施用尿素的处理相比，施用控释氮肥后，早、晚稻生育中、后期功能叶中的叶绿素含量得到显著的提高，且与尿素间的差异随着水稻生育进程而增大。不同时期控释氮肥处理功能叶的净光合速率也均显著地高于尿素处理。此外，施用控释氮肥能显著地提高生育中、后期功能叶中的SOD、POD等活性氧清除酶的活性，降低功能叶中的丙二醛含量。施用控释氮肥对功能叶中的IAA、ABA含量具有明显的调控效果，早、晚稻各时期控释氮肥处理的IAA含量均显著地高于尿素处理，而ABA含量显著地低于尿素处理。由于控释氮肥对水稻的上述调控机理，因而，在水稻上施用控释氮肥可以明显提高水稻产量。     

Morphological and physiological traits of seeds and seedlings in two rice cultivars with contrasting early vigor

Early vigor is important for crop establishment in rice. This study was conducted to determine the seed and seedling traits in relation to early vigor in rice. Laboratory tests and pot experiments were carried out in 2013 and 2014. Morphological and physiological traits of seeds and seedlings were compared between two contrasting rice cultivars, Yuxiangyouzhan with superior early vigor and Huanghuazhan with general early vigor. For seed traits, Yuxiangyouzhan had lower seed hull weight but higher seed amylose content, seed amylase activity, and plumule–radicle ratio than Huanghuazhan, and consequently, Yuxiangyouzhan had about 10% higher germination percentage and velocity and 37% longer plumule than Huanghuazhan. For seedling traits, leaf area, specific leaf weight, leaf chlorophyll and soluble protein contents, leaf net photosynthetic rate, and shoot–root ratio were higher in Yuxiangyouzhan than in Huanghuazhan, and as a result, seedling shoot biomass was 23–32% higher in Yuxiangyouzhan than in Huanghuazhan. These results indicate that Yuxiangyouzhan has both superior seed and seedling vigor. The former is attributed to the low mechanical strength of seed hull, high conversion efficiency of seed reserve, and high mobilization of seed reserve to plumule, while the latter is due to large leaf area, high leaf photosynthetic capacity, and high partitioning of dry matter to shoot. This study enriches the physiological understanding of superior early vigor in rice.     

Effects of GS3 and GL3.1 for Grain Size Editing by CRISPR/Cas9 in Rice

Grain size is one of key agronomic traits associated with grain yield and grain quality. Both major quantitative trait loci GS3 and GL3.1 play a predominant role in negative regulation of grain size. In this study, a CRISPR/Cas9-mediated multiplex genome editing system was used to simultaneously edit GS3 and GL3.1 in a typical japonica rice Nipponbare. In T_1 generation, we found that gs3 formed slender grain with lower chalkiness percentage, while gs3gl3.1 produced larger grain with higher chalkiness percentage. In terms of other agronomic traits, flag leaf size, grain number and grain yield of both gs3 and gs3gl3.1 mutants were affected. It is noteworthy that gs3 and gs3gl3.1 mutants both led to dramatical reduction of grain number, thereby decreased grain yield. In conclusion, these results indicated that knockout of GS3 and GL3.1 could rapidly improve grain size, but probably have some negative influences on grain quality and grain yield.     

利用CRISPR/Cas9系统定向改良水稻粒长和穗粒数性状

【目的】基因组定点编辑技术已成为分子育种的重要手段。本研究拟对GS3和Gn1a功能缺失突变对目标性状的改良效应进行分析,以期为培育高产水稻提供理论基础。【方法】利用CRISPR/Cas9系统,以控制粒型基因GS3和控制每穗粒数基因Gn1a为编辑对象,构建了共敲除载体p C1300-2×35S::Cas9-g~(GS3)-g~(Gn1a)a,用农杆菌介导法转化4个优质水稻品种,分析了基因突变的特征和相应农艺性状。【结果】构建的敲除载体成功地实现了对GS3和Gn1a基因的定点编辑。在4个转化受体的T_0代均分别获得了gs3和gs3gn1a的移码突变体。对T_1 代中无选择标记突变体的农艺性状分析表明,突变体gs3和gs3gn1a与野生型相比粒长变长,千粒重增加;突变体gs3gn1a与突变体gs3相比,每穗粒数显著增加。【结论】利用CRISPR/Cas9系统进行水稻基因编辑可以快速改良品种的目标性状,在水稻品种的定向改良方面具有巨大的潜力。     

EMS诱导的盛农1号小麦突变体筛选与鉴定

突变体诱导是创造小麦遗传材料和选育新品种的重要途径之一。通过甲基磺酸乙酯（EMS）处理高产、抗白粉病品系盛农1号,从4 500个M₂代穗系中筛选出174个突变体穗系,突变率为3.86%。其中,穗部突变体32个,突变率为0.71%;叶部突变体47个,突变率为0.89%;茎部突变体22个,突变率为0.49%;育性和成熟期突变体12个,突变率为0.27%;其他类型突变体11个,突变率为0.24%;农艺性状优于对照盛农1号的突变体57个,突变率为1.27%。经过M₃代验证及生物学性状和农艺性状调查,获得可稳定遗传的株系18个,其中,穗部突变体6个,茎秆突变体1个,叶部突变体6个,早熟突变体3个,晚熟突变体2个。农艺性状分析表明,穗部突变体SS4、SS5、SS6和SS16的小穗性状优于盛农1号。这些新突变体为小麦品种改良和遗传研究提供了新的材料。     

水稻水作与旱作对抽穗后剑叶光合特性、衰老及根系活性的影响

对水稻水作与旱作栽培模式下水稻衰老和后期一些生理活性的影响进行了研究。结果表明,由于水分的影响,水稻旱作不同程度加速了水稻生理机能的衰退,使籽粒干物质积累受阻,粒重下降而减产。水稻水作不仅改善了水稻剑叶光合因子,提高了光合效率,而且剑叶呼吸速率下降缓慢,相对电导率上升缓慢,SOD酶活性较高,MDA含量低,表明剑叶后期衰老进程慢,促进籽粒增重,产量高。此外,旱作水稻生育末期根系干物质积累、活力高及代谢旺盛的优势对籽粒增重也是一种负向影响;而水稻水作开花-灌浆期根系活力高,生育末期根系生理活性迅速衰退,保证了籽粒的充实