东北中部地区水稻不同生育时期低温处理下生理变化及耐冷性比较

摘要：研究水稻不同生育时期低温条件下各种生理指标的变化,选育耐低温水稻新品种,对于东北地区的水稻生产具有重要意义。以东北中部地区目前主要栽培的15个主要栽培水稻品种为实验材料,利用人工气候箱进行低温处理,分析苗期、孕穗期、抽穗期和灌浆期4个不同生育时期各个品种过氧化物酶（POD）、丙二醛（MDA）、脯氨酸和叶绿素4种生理指标含量变化与耐低温系数之间的关系。同时利用不同生理指标隶属函数的加权平均值（D值）对不同水稻品种的抗冷性进行分类划分。结果表明,每个生育时期的低温处理下,过氧化物酶（POD）、脯氨酸、叶绿素均呈下降趋势,丙二醛（MDA）则呈上升趋势,并且4种指标的变化幅度与水稻耐低温系数之间呈极显著相关性（P＜0.01）,各种指标的加权平均值（D值）与抗低温系数之间相关系数达到极显著,并且高于每一个单一指标。因此,D值可以更有效的对水稻进行抗低温评价和分级。     

低温胁迫下粳稻选育品种耐冷性状的鉴定评价

选取来源于中国11个省份和其他9个国家的347份粳稻选育品种作为试验材料,分析了自然低温和冷水胁迫下,不同来源粳稻选育品种孕穗期的耐冷性及主要农艺性状的表型差异和聚类特点。研究表明,在自然低温和冷水胁迫下各省份或国家粳稻选育品种主要农艺性状及其冷水反应指数有明显的差异。在自然低温和冷水胁迫下,云南和日本品种的孕穗期结实率及其冷水反应指数均较高,表现出较强的孕穗期耐冷性。从总体趋势上看,在自然低温下,除个别省份外,我国纬度相对较高的北方省份品种的孕穗期耐冷性强于纬度相对较低的南方省份品种;而在冷水胁迫下,品种的耐冷性与其来源地的关系并不密切,没有呈现出一定的规律性。此外,聚类结果表明,不同省份或国家粳稻选育品种的聚类结果与其品种的地理来源均有一定的相关性,而与自然条件相比,冷水胁迫下粳稻选育品种的聚类结果与其品种的地理来源的相关性更为密切。     

中国水稻微核心种质不同生育时期耐冷性鉴定及其相关分析

以204份中国水稻微核心种质为试验材料,进行了水稻发芽期、芽期、幼苗期、孕穗期等耐冷性鉴定及其相关性分析。结果表明,水稻各生育时期耐冷性在籼粳亚种间、各种质之间存在明显的差异。粳稻种质的耐冷性明显强于籼稻种质,但籼稻种质中也存在耐冷性较强的种质。高阳淀稻大红芒、肥东塘稻、木樨球、卫国、兴国、山酒谷、中花8号等粳稻种质和包选21号、红米三担白、寸谷糯、红谷等籼稻种质在水稻各生育时期均表现较强的耐冷性,在水稻耐冷性育种及耐冷基因发掘研究中应加以利用。自然低温和冷水胁迫下水稻结实率与低温下发芽率呈显著或极显著正相关,而与死苗率呈显著负相关,即低温下发芽期耐冷性和芽期耐冷性强的水稻种质一般表现为较强的孕穗期耐冷性。认为低温下发芽率和芽期低温处理后的死苗率可以作为孕穗期耐冷性早期鉴定的间接指标。     

中国水稻微核心种质不同生育时期耐冷性鉴定及其相关分析

以204份中国水稻微核心种质为试验材料,进行了水稻发芽期、芽期、幼苗期、孕穗期等耐冷性鉴定及其相关性分析。结果表明,水稻各生育时期耐冷性在籼粳亚种间、各种质之间存在明显的差异。粳稻种质的耐冷性明显强于籼稻种质,但籼稻种质中也存在耐冷性较强的种质。高阳淀稻大红芒、肥东塘稻、木樨球、卫国、兴国、山酒谷、中花8号等粳稻种质和包选21号、红米三担白、寸谷糯、红谷等籼稻种质在水稻各生育时期均表现较强的耐冷性,在水稻耐冷性育种及耐冷基因发掘研究中应加以利用。自然低温和冷水胁迫下水稻结实率与低温下发芽率呈显著或极显著正相关,而与死苗率呈显著负相关,即低温下发芽期耐冷性和芽期耐冷性强的水稻种质一般表现为较强的孕穗期耐冷性。认为低温下发芽率和芽期低温处理后的死苗率可以作为孕穗期耐冷性早期鉴定的间接指标。     

Effect of Chilling on Photosynthesis in Flag Leaves at Primary Heading Stage of F1 Hybrid Rice Cultivars and Their Parental Lines (Three Lines)

With the purpose of selecting and breeding cold stress tolerance of hybrid rice cultivars, the effect of chilling on five hybrid rice combinations was investigated. The results indicated that the cold tolerance of different hybrid rice varieties were different. The order of the cold tolerance as the degrees of inhibition of maximum photosynthetic rate and of apparent quantum yield of flag leaves at primary heading stage by chilling was as follows: The cold tolerance of Japanica type Xiuyou-57 was the highest then lndica type Qing-Youzao followed by Shangyou-63, Shangyou-64 and Weiyou-64. The cold tolerance of Fx hybrid rice was similar to that of the maternal lines and not similar to the paternal lines. The cold tolerance of maintainer lines was similar to that of the male sterile lines. The tests on the survival rate of hybrid rice seedlings after chilling treatment also showed a similar re gular patterns. These indicated that the responses by the chilling temperature in these five hybrid combinations were exactly the same in the Seedling stage as well as in the late growing stage. The results of these experiments on the impact of selecting and breeding cold tolerance in hybrid rice were discussed.     

孕穗期低温对黑龙江省主栽水稻品种空壳率的影响

障碍型冷害对黑龙江省水稻生产影响巨大,明确主栽水稻品种耐冷性及障碍型冷害的量化指标,可为当地水稻安全生产提供科学依据。本文对黑龙江省4个主栽水稻品种(垦稻12、龙稻3号、龙稻7号和空育131)在孕穗期进行不同低温处理,分析了各处理对水稻空壳率的影响。结果表明:在平均温度为15℃时,孕穗期低温的持续天数是水稻形成障碍型冷害的关键因素;在低温持续时间相同的条件下,昼夜恒温15℃使各品种水稻受低温影响最严重;在不同低温处理条件下,不同品种水稻发生一般障碍型冷害的临界时长为1～4d或5～8d,发生严重障碍型冷害的临界时长为5～8d或8d以上,品种间临界时长有所差异;供试品种中,垦稻12和龙稻3号耐冷性较弱,空育131耐冷性较强,龙稻7号耐冷性居中。     

低温对杂交水稻及其亲本三系始穗期旗叶光合作用的影响

研究了低温对5套杂交水稻组合的影响。始穗期旗叶光合作用的最大速率和表观量子效率受低温抑制的程度表明:其耐冷顺序为粳稻型的“秀优57”>“釉稻型的青优早”>“汕优63”、“汕优64”与“威优64”。杂交水稻 F_1的耐冷水平与母本近似,与父本关系不大,保持系的耐冷性相似于不育系。杂交水稻幼苗经低温处理后的存活率亦表现出同样的规律。证明所试5套杂交水稻的生育后期与苗期对低温的反应是一致的。本文对该结果在杂交水稻抗冷性选育种中的意义进行了讨论。     

广西水稻地方品种耐冷性鉴定及相关分析

对419份广西水稻地方品种初级核心种质进行芽期、苗期的耐冷性鉴定及相关分析,结果表明:广西水稻地方品种芽期、苗期耐冷性主要集中在7级和9级,总体耐冷性较弱。芽期、苗期极强耐冷种质(1级)分别为24份和27份,占参试总数的5.73%和6.44%,其中10份种质芽期和苗期均表现极强耐冷(1级)。芽期、苗期耐冷性呈极显著正相关(r=0.66)。粳稻芽期、苗期耐冷性均显著高于籼稻;粘糯稻之间耐冷性差异是由籼粳稻类型的耐冷差异引起的;来自高寒山区稻作区的品种芽期和苗期平均耐冷表现最强。利用34个SSR标记与芽期、苗期耐冷性进行Pearson相关分析,在第7和第9染色体上,各鉴定出1个同时与芽期和苗期耐冷性相关联的位点。本研究为水稻芽期、苗期耐冷育种提供新的抗源材料,并为水稻耐冷基因定位及机理研究奠定基础。     

Genetic Analysis of Cold Tolerance at the Germination and Booting Stages in Rice by Association Mapping

Low temperature affects the rice plants at all stages of growth. It can cause severe seedling injury and male sterility resulting in severe yield losses. Using a mini core collection of 174 Chinese rice accessions and 273 SSR markers we investigated cold tolerance at the germination and booting stages, as well as the underlying genetic bases, by association mapping. Two distinct populations, corresponding to subspecies indica and japonica showed evident differences in cold tolerance and its genetic basis. Both subspecies were sensitive to cold stress at both growth stages. However, japonica was more tolerant than indica at all stages as measured by seedling survival and seed setting. There was a low correlation in cold tolerance between the germination and booting stages. Fifty one quantitative trait loci (QTLs) for cold tolerance were dispersed across all 12 chromosomes; 22 detected at the germination stage and 33 at the booting stage. Eight QTLs were identified by at least two of four measures. About 46% of the QTLs represented new loci. The only QTL shared between indica and japonica for the same measure was qLTSSvR6-2 for SSvR. This implied a complicated mechanism of old tolerance between the two subspecies. According to the relative genotypic effect (RGE) of each genotype for each QTL, we detected 18 positive genotypes and 21 negative genotypes in indica, and 19 positive genotypes and 24 negative genotypes in japonica. In general, the negative effects were much stronger than the positive effects in both subspecies. Markers for QTL with positive effects in one subspecies were shown to be effective for selection of cold tolerance in that subspecies, but not in the other subspecies. QTL with strong negative effects on cold tolerance should be avoided during MAS breeding so as to not cancel the effect of favorable QTL at other loci.     

Effects of Chilling Temperature on Photosynthesis and Photosynthate Transport in Flag Leaves of Hybrid Rice at Milky Stage

To understand the physiological reasons for poor yield of the second rice crop in southern China challenged by low temperature. The authors investigated the effects of chilling temperature on photosynthesis and the activity of fructose 1, 6-diphosphatase (FBPase) of flag leaves at milky stage using two hybrid rice varieties with different cold tolerance. The results indicated that chilling temperature caused decreases of photosynthetic efficiency and FBPase activity in detached flag leaves. The decline of photosynthetic efficiency and FBPase activity was greater in Shanyou-63 which is as cold sensitive as its parents than in Xiuyou-57 which is cold tolerant like its parents. The milky stage is the period of the fastest grain filling. The decreased the yield of the second rice crop caused by natural low temperature was associated with decline of photosynthesis and the abilities of adaption and adjustment of FBPase in attached flag leaves. The relationship between the effects of chilling temperature on photosynthesis and photosynthate transport and the yield formed by grain was discussed.     

低温对杂交水稻乳熟期剑叶光合作用和光合产物运输的影响

为了解寒露风引起晚稻减产的生理原因,本试验以两个不同抗冷力的杂交水稻组合为材料,研究了低温对水稻乳熟期剑叶光合作用与果糖１,６－二磷酸酯酶（ＦＢＰａｓｅ）活性的影响。结果表明：低温引起离体剑叶光合效率与ＦＢＰａｓｅ 活性下降,不抗冷的“汕优６３”及其亲本的光合效率与ＦＢＰａｓｅ 活性比抗冷的“秀优５７”及其亲本下降幅度较大。乳熟期是籽粒灌浆的高峰期,自然低温（寒露风）造成晚造水稻减产与乳熟期连体剑叶的光合作用下降和ＦＢＰａｓｅ 对低温的适应和调节能力有关     

黑龙江省水稻空壳率与孕穗期低温的关系

对黑龙江省6个主要水稻品种(龙稻3号、垦稻12号、空育131、龙稻7号、龙粳16号和松粳6号)进行孕穗期低温(处理温度分别为15℃、17℃、19℃,低温持续时间分别为2、4、6和8d)处理,采用线性内插和统计回归方法,分析了水稻单穗空壳率与孕穗期低温的关系.结果表明:研究区敏感性水稻品种和耐冷性较强水稻品种的障碍型冷害临界温度分别为17℃和16℃;孕穗期水稻对低温最敏感的时期为抽穗前14～18d.15℃低温处理8d时,松粳6号、垦稻12号的空壳率明显增加,空育131空壳率小幅升高,表明松粳6号和垦稻12号对低温反应较敏感、耐冷性较弱,而空育131对低温反应迟钝,耐冷性较强;垦稻12号、龙粳16号、空育131的冷积温与空壳率存在显著的相关关系(P0.01),随着冷积温的增加,水稻空壳率明显升高,但品种间的增幅不同.     

Proteomic Analysis of Rice Seedlings Under Cold Stress

Low temperature can greatly restrict the growth and development of rice. The rice seedlings show growth retardation, lamina wrap, and part of blade even died under the condition of low temperature. In order to get more information about cold stress responses in rice, two dimensional electrophoresis and bioinformatics analysis of mass spectrometry were used to preliminary survey the cold tolerance of cold sensitive line 9311 and cold resistance variety Fujisaka 5 under cold stress. Two dimensional electrophoresis maps of 9311 and Fujisaka 5 were established under cold treatment. With analysis of bioinformation, the proteins were found involve in many aspects of rice development. The largest category of proteins is functioning on metabolism. By comparing the proteins from the two varieties, it can be found that most proteins from 9311 were down-regulated and were up-regulated in Fujisaka 5. The results showed that the membrane composition and structure were damaged, metabolism changed dramatically and rice defense system was activated under the cold stimulation. Fifty-nine proteins related to the resistance of cold stress were identified in our study, and we have investigated and classified all of their biological functions. The importance of our study are providing some conduct for the research of rice resistant to cold stress, supporting auxiliary technique for rice varieties and widening the search field of cold tolerance in plants.     

Effects of Shading on Starch Pasting Characteristics of Indica Hybrid Rice (Oryza sativa L.)

Rice is an important staple crop throughout the world, but environmental stress like low-light conditions can negatively impact crop yield and quality. Using pot experiments and field experiments, we studied the effects of shading on starch pasting viscosity and starch content with six rice varieties for three years, using the Rapid Visco Analyser to measure starch pasting viscosity. Shading at different growth stages and in different rice varieties all affected the starch pasting characteristics of rice. The effects of shading on starch pasting viscosity at middle and later growth stages were greater than those at earlier stages. Shading enhanced breakdown but reduced hold viscosity and setback at tillering-elongation stage. Most pasting parameters changed significantly with shading after elongation stage. Furthermore, the responses of different varieties to shading differed markedly. The change scope of starch pasting viscosity in Dexiang 4103 was rather small after heading, while that in IIyou 498 and Gangyou 906 was small before heading. We observed clear tendencies in peak viscosity, breakdown, and pasting temperature of the five rice varieties with shading in 2010 and 2011. Correlation analysis indicated that the rice amylose content was negatively correlated with breakdown, but was positively correlated with setback. Based on our results, IIyou 498, Gangyou 906, and Dexiang 4103 had higher shade endurance, making these varieties most suitable for high-quality rice cultivation in low-light regions.     

中国粳稻地方品种孕穗期耐冷性评价及聚类分析

用原产于中国18个省的329份粳稻地方品种为材料,分析了自然低温和冷水胁迫下不同省份粳稻地方品种孕穗期耐冷性及主要农艺性状表型差异和聚类特点。结果表明,在自然低温和冷水胁迫下各省份粳稻地方品种的孕穗期耐冷性状及主要农艺性状有明显的差异。在自然低温和冷水胁迫下,天津、四川和台湾品种的结实率及冷水反应指数均较高,表现较强的孕穗期耐冷性和迟钝的冷水反应。云南品种在自然低温下表现为较强的孕穗期耐冷性（结实率）,而在冷水胁迫下播种至抽穗天数和株高的冷水反应指数较高,表现为迟钝的冷水反应。从总体趋势上看,自然低温和冷水胁迫下,除个别省份外,纬度相对较高的北方省份品种的孕穗期耐冷性（结实率）强于纬度相对较低的南方省份品种。此外,在自然低温和冷水胁迫下,各省粳稻地方品种的聚类结果总体上与各省品种的地理位置及其耐冷性有密切的联系,而在冷水胁迫下品种的聚类结果与品种地理位置的关系比自然低温下更为密切。     

Mapping of Quantitative Trait Loci Underlying Cold Tolerance in Rice Seedlings via High-Throughput Sequencing of Pooled Extremes

Low temperature is a major limiting factor in rice growth and development. Mapping of quantitative trait loci (QTLs) controlling cold tolerance is important for rice breeding. Recent studies have suggested that bulked segregant analysis (BSA) combined with next-generation sequencing (NGS) can be an efficient and cost-effective way for QTL mapping. In this study, we employed NGS-assisted BSA to map QTLs conferring cold tolerance at the seedling stage in rice. By deep sequencing of a pair of large DNA pools acquired from a very large F₃ population (10,800 individuals), we obtained ∼450,000 single nucleotide polymorphisms (SNPs) after strict screening. We employed two statistical methods for QTL analysis based on these SNPs, which yielded consistent results. Six QTLs were mapped on chromosomes 1, 2, 5, 8 and 10. The three most significant QTLs on chromosomes 1, 2 and 8 were validated by comparison with previous studies. Two QTLs on chromosomes 2 and 5 were also identified previously, but at the booting stage rather than the seedling stage, suggesting that some QTLs may function at different developmental stages, which would be useful for cold tolerance breeding in rice. Compared with previously reported QTL mapping studies for cold tolerance in rice based on the traditional approaches, the results of this study demonstrated the advantages of NGS-assisted BSA in both efficiency and statistical power.     

低温胁迫下不同甘蔗品种(系)光合特性的变化及其与耐寒性的关系

以广西农科院甘蔗研究所自育的7个新材料和2个生产上的主栽品种为研究对象,在甘蔗苗期进行低温胁迫处理,研究了各品种(系)甘蔗形态特征的冷害指数、叶绿素含量及光合特性相关指标的变化及其光合特性相关指标与甘蔗抗寒性间的相关性。结果表明:随着低温胁迫处理时间的延长,冷害指数不断增大,但变化的大小与快慢因品种(系)不同表现不一样。各甘蔗品种(系)叶片叶绿素含量均随时间延长而降低。叶片净光合速率、气孔导度在低温处理与常温处理间具有显著差异。低温胁迫处理显著降低了各甘蔗品种(系)最大光化学效率(Fv/Fm)、PSⅡ实际光能转化效率ΦPSⅡ、光适应下PSⅡ反应中心的最大光能转化效率Fv′/Fm′、光化学猝灭系数qP、电子传递速率ETR,而显著提高了初始荧光Fo、稳态荧光Fs、非光化学猝灭系数qNP。相关性分析表明整个测定时期各指标间相关显著,Fv/Fm、Fv′/Fm′、ΦPSⅡ与冷害指数I之间的相关系数在0.800以上,Fv/Fm、Fv′/Fm′、ΦPSⅡ可以作为甘蔗品种(系)抗寒性鉴定的重要参考指标。     

A point mutation in LTT1 enhances cold tolerance at the booting stage in rice

The cold tolerance of rice at the booting stage is a main factor determining sustainability and regional adaptability. However, relatively few cold tolerance genes have been identified that can be effectively used in breeding programmes. Here, we show that a point mutation in the low-temperature tolerance 1 (LTT1) gene improves cold tolerance by maintaining tapetum degradation and pollen development, by activation of systems that metabolize reactive oxygen species (ROS). Cold-induced ROS accumulation is therefore prevented in the anthers of the ltt1 mutants allowing correct development. In contrast, exposure to cold stress dramatically increases ROS accumulation in the wild type anthers, together with the expression of genes encoding proteins associated with programmed cell death and with the accelerated degradation of the tapetum that ultimately leads to pollen abortion. These results demonstrate that appropriate ROS management is critical for the cold tolerance of rice at the booting stage. Hence, the ltt1 mutation can significantly improve the seed setting ability of cold-sensitive rice varieties under low-temperature stress conditions, with little yield penalty under optimal temperature conditions. This study highlights the importance of a valuable genetic resource that may be applied in rice breeding programmes to enhance cold tolerance.     

陆地棉萌发至三叶期不同生育阶段耐盐特性

摘要：土壤盐渍化是一个世界性的资源问题和生态问题,盐分胁迫几乎会影响棉花所有重要的生命活动, 造成棉花的减产或其他不利影响。本试验通过对14个品棉花种（系）的萌发期耐盐性、芽期耐盐性以及两个耐盐性不同的品种（系）三叶期的耐盐性,对棉花不同时期的耐盐特性进行了研究,结果表明：棉花在不同生育阶段耐盐能力是不同的,棉花盐敏感时间是在萌发出苗期。随着生育期的延长,棉花的耐盐性是逐步提高的,进行耐盐性鉴定的最佳时期在是萌发出苗期。在三叶期,随着盐分浓度的提高,棉花苗受伤害程度逐渐增加,盐敏感性品种受伤害大于耐盐品种,在同样盐浓度胁迫条件下,耐盐性不同的品种各部分受到的伤害不同,耐盐品种（系）中棉所35受伤害程度由高到低依次为：真叶>上胚轴>子叶;盐敏感品种（系）Tamcot CAB-CS受伤害程度由高到低依次为：真叶> 子叶>上胚轴。     

Identification and pyramiding of QTLs for cold tolerance at the bud bursting and the seedling stages by use of single segment substitution lines in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

Cold stress is one of the main constraints in rice production, and damage from cold can occur at different developmental stages in rice. Understanding the genetic basis of cold tolerance is the key for breeding cold-tolerant variety. In this study, we used single segment substitution lines (SSSLs) derived from a cross between cold-tolerant japonica variety “Nan-yang-zhan” and a popular indica variety “Hua-jing-xian 74” to detect and pyramid QTLs for cold tolerance at the bud bursting and the seedling stages. Evaluation of cold tolerance of these SSSLs and their recurrent parent helped identify two cold-tolerant QTLs (qCTBB-5 and qCTBB-6) at the bud bursting stage and two cold-tolerant QTLs (qCTS-6 and qCTS-12) at the seedling stage. The SSSLs carrying these QTLs showed stronger cold tolerance than their recurrent parent HJX74 did in three independent experiments. The qCTBB-6 and qCTS-6 were mapped to the same chromosomal region. QTL pyramiding was performed by intercrossing of SSSLs carrying the respective QTLs for cold tolerance at the bud bursting stage and the seedling stage and marker-assisted selection (MAS). The selected pyramiding line SC1-1 with different cold-tolerant QTLs showed cumulative effects on cold tolerance. Our results suggest that different genes (QTLs) control cold tolerance at bud bursting and seedling stages, and pyramiding of stable expression QTLs for cold tolerance at different developmental stages through MAS is a good strategy to prevent cold damage in rice.