应用近等基因系初步定位粳稻孕穗期的耐冷基因

 在昆明低温冷害条件下对十和田×(十和田5/昆明小白谷BC4F5)的BC5F2群体进行了耐冷性状的遗传研究。结果显示，穗期耐冷性受贡献率较大的基因控制。用平均分布于12条染色体的164个SSR标记对十和田、昆明小白谷、近等基因系池（NILP）进行筛选，在第5染色体长臂末端有2个SSR标记的扩增产物在十和田与昆明小白谷、NILP间有多态性。用这2个多态性标记对群体进行分子标记定位，单向方差分析表明RM31与耐冷基因连锁。再在RM31附近合成12个SSR标记在轮回亲本（RP）、近等基因系（NIL）间进行多态性筛选，只有RM7452有多态性, 单向方差分析表明该标记与耐冷基因连锁。耐冷基因与RM7452、RM31的遗传距离分别为4.8 cM和8.0 cM，主穗结实率能解释表型变异的10.50%；实粒数能解释表型变异的5.10%。暂将这个耐冷基因定名为Ctb(t)。     

粳稻02428×02428c重组自交系孕穗期耐冷性QTL分析

 利用孕穗期弱耐冷性品种02428和强耐冷性品种02428c构建了一个包含336个株系的F5：6重组自交系群体。采用自然低温冷害鉴定法,以实粒数、秕粒数、总粒数和结实率作为孕穗期耐冷性状的表型值,用486对SSR标记初步构建分子连锁图谱,对孕穗期耐冷性进行QTL定位。结果表明,标记RM6092、RM6702、RM5954、RM1095、RM1183、RM7643及RM3411与孕穗期耐冷性状连锁;利用QTLMapper 1.6软件检测到位于第1染色体上的3个耐冷性QTL位点。     

普通野生稻苗期耐冷性QTL的鉴定与分子定位

 以两份普通野生稻核心种质资源DP15和DP30为供体、9311为受体构建染色体片段代换系鉴定苗期耐冷性QTL;利用苗期耐冷性最强的1个代换系构建QTL作图群体,用SSR标记对其主效QTL进行定位。研究结果表明,两个抗源DP15和DP30所含的苗期耐冷性QTL的数量、位点及耐冷性效应均存在明显的差异。在基本上覆盖两个亲本全基因组的230份BC4F2代换系中共发现19个苗期耐冷性QTL,分布在水稻12条染色体上,第3和第8染色体上有比较密集的苗期耐冷性QTL分布。这19个分布于全基因组的苗期耐冷性QTL被分别分离到不同的野生稻染色体片段代换系里,效应最小的微效QTL位点所在的代换系在苗期耐冷性鉴定中的活苗率仅为8%,而效应最大的主效QTL位点所在代换系的活苗率达到74%。这个主效QTL qSCT 3 1被定位在第3染色体着丝点附近长臂上的RM15031―RM3400区间,距离最近的标记RM15040、RM1164的遗传距离为1.8 cM。     

籼粳稻区云南稻种耐冷性状遗传变异研究

 用5个不同耐冷级别品种按1/2p（p-1）配制15个组合及其亲本在籼稻区（F1）、籼粳交错区（F2）和温凉粳稻区（F1、F2、F3）进行云南稻种颖壳、花药大小和结实率的遗传变异研究,结果表明：（1）耐冷稻种在冷害条件下颖壳变窄、花药大小相对稳定、结实率较高,花药长缩短与结实率呈负相关或极显著负相关; 极弱耐冷品种十和田则呈极显著正相关(0.924**)。（2）极强耐冷的昆明小白谷、丽粳2号耐冷性状的GCA较高,用它们作耐冷基因供体、十和田作轮回亲本已获得一批耐冷性近等基因系。（3）不同世代同一稻区和同世代不同稻区的耐冷性状及SCA在不同稻区存在明显差异。（4）籼粳杂种后代在温凉粳稻区粳型性状变异同时伴随着耐冷基因的累积;而在籼粳交错区和籼稻区则有利于籼型性状的遗传进化。     

水稻冷胁迫响应基因OsSADMC功能标记的开发和利用

通过对95份水稻种质资源进行苗期耐冷性鉴定,筛选出10份苗期耐冷(存活率≥60%)和9份冷敏感的材料(存活率为零),并对水稻冷胁迫响应基因OsSADMC进行了表达分析,发现对照和冷胁迫处理条件下该基因在耐冷品种中的表达量都显著高于冷敏感品种。对耐冷材料丽江新团黑谷(LTH)和冷敏感材料IR29的基因编码区域进行了克隆和测序分析,在两个品种间鉴定了10个SNP位点,其中第749碱基处存在的一个碱基变异(C突变为T),从而使丝氨酸变为亮氨酸。根据其中一个SNP位点设计了OsSADMC的功能标记SADMC-CAPS1,该标记可以准确地鉴定出19份耐冷性不同水稻种质资源OsSADMC的基因型和耐冷性。     

粳稻体细胞耐冷变异的诱导筛选技术

为建立水稻耐冷育种的新途径,开展了本项试验研究。结果是：（１）在１５℃下愈伤组织净生长量与供体品种孕穗期的耐冷性呈的正相关。其中２５天的相关系数最大。（２）１５℃下选择的愈伤组织的无性系中６２．５％的无性系耐冷性明显强于供体品种;（３）体细胞无性系２,３代的广义遗传力高。（４）供体品种孕穗期的耐冷性与耐冷变异性体率呈极显著的负相关,与感冷变异体率呈正相关。     

水稻幼苗期耐冷性选择对主要农艺性状的影响

利用籼粳稻杂交后代（密阳23/通88-7、密阳23/TR22183、密阳23/高产102）探讨了水稻幼苗期耐冷性选择对主要农艺性状的影响。从籼粳稻杂交后代F2和F3中选择的耐冷个体群或系统群与随机个体群或敏冷系统群间进行主要农艺性状的比较结果表明,在F2和F3水稻幼苗期耐冷性选择对秆长、穗长和穗数并不产生显著的影响,但选择获得的幼苗期耐冷性强的F2个体群的孕穗期耐冷性(结实率)、F3系统群的分蘖期耐冷性(赤枯度)和孕穗期耐冷性(结实率)均显著强于随机个体群和敏冷系统群;在冷水处理下F3幼苗期耐冷性强的耐冷系统群的产量显著高于敏冷系统群。提出在水稻幼苗期以叶赤枯度来选择幼苗期耐冷性强的个体是获得高产耐冷后代材料的有效途径。     

大豆萌发期对6℃低温的反应

1987—1988年,利用人工气候箱对我国东北春大豆品种(系)1910份进行了萌发期抗冷的筛选与研究,温度控制于6℃,恒温,并以25℃发芽实验作对照。(1)不同品种(系)在6℃下发芽率变化很大,其范围从0—100％;(2)黑龙江大豆在6℃下发芽速率最快;(3)确定了6℃下发芽时的最佳调查时间为13—14天;(4)两年共筛选出358个高抗冷材料,占参试材料的18.7％;(5)黑种皮,深褐脐,肾状、扁椭粒,种皮无光泽表现出较强的抗冷性;(6)百粒重与抗冷呈极显著的负相关;(7)种子的蛋白质含量与抗冷性关系不大,粗脂肪含量、脯氨酸含量与抗冷性呈极密切的负相关,亚油酸与油酸的比值与抗冷性呈极显著的正相关。     

水稻种子含水量与耐冷性的关系研究

试验选择18份水稻品种的休眠种子作为材料,其中具有耐冷性的水稻品种13份,不具有耐冷性的常规水稻品种5份。测定了每份种子样品中含水量,并将这项指标分别与水稻品种是否具有耐冷性以及耐冷性强弱进行相关性分析。结果表明：①水稻品种是否具有耐冷性与种子中含水量呈极显著负相关关系,即耐冷性品种含水量明显低于常规品种;②具有耐冷性的水稻品种,其耐冷性程度与种子的含水量没有显著的相关性。因此认为水稻休眠种子的含水量可以作为判断水稻耐冷性有无的参考指标,但不能直接用作判定水稻耐冷性强弱的指标。     

利用关联分析方法挖掘自然群体中大豆油分和蛋白质含量相关SSR标记

为研究自然群体中大豆品种的油分和蛋白质含量变化,筛选出相关标记的优异等位变异,以327份东北主推品种和优异亲本材料构成的自然群体为试验材料。以分布于大豆20条染色体的186对SSR(simple sequence repeat)分子标记检测所有试验材料的基因型,利用STRCTURE 2.3.4软件分析群体结构,将试验材料分为7个亚群。利用TASSEL 3.0软件的混合线性模型的方法对大豆自然群体的油分和蛋白质含量进行关联分析。在极显著水平(P0.01)且贡献率大于1%情况下,共检测到33个关联位点。与油分含量极显著关联位点12个,解释率为2.19%~10.05%;与蛋白质含量极显著关联位点11个,解释率为2.65%~9.08%;与油分和蛋白质含量同时关联位点6个,分别为Satt005、Satt117、Satt565、Satt469、Satt594和Satt546,其中Satt594解释率最高,油分为10.05%,蛋白质为9.08%。     

水稻生长早期耐冷性QTL分析

 以籼粳交“密阳23/吉冷1号”的F2：3代200个家系为作图群体，构建分子连锁图谱，并进行了F3代家系的生长早期耐冷性鉴定和QTL分析。结果表明，幼苗期耐冷性、分蘖期耐冷性和低温下幼苗生长能力等生长早期耐冷性在F3代表现为近似正态的连续分布，是由多基因控制的数量性状。在第1、5、9染色体上分别检测到与幼苗期耐冷性相关的QTL各1个，其中qCTS1对表型变异的解释率最大，达15.5%；在第2、3、7、9、11染色体上分别检测到与分蘖期耐冷性相关的QTL各1个，而每个QTL对表型变异的解释率均较低；在第1、2、11、12染色体上分别检测到与低温下幼苗生长能力相关的QTL各1个，其中qGAS2和qGAS12对表型变异的解释率分别为26.6%和42.9%，是主效基因。     

Mapping a Novel Gene of Cold Tolerance at Booting Stage by Using Near-Isogenic Lines in japonica Rice

Genetic analysis showed that cold tolerance at booting stage of near-isogenic lines （NILs） of Kunmingxiaobaigu was controlled by a gene with large phenotypic variance. One hundred and sixty-four simple sequence repeats （SSR） distributed over 12 chromosomes were used to screen polymorphism between Towata （recurrent parent, RP） and near-isogenic line pool （NILP）, and two SSR markers at the long arm of chromosome 5 showed polymorphism in comparison with RP genome. Of the two markers, RM31 was found possibly linked with the cold tolerance gene at booting stage through one-way ANOVA. Twelve SSR markers around RM31 were then used to detect polymorphism between RP and NIL, and only RM7452 had polymorphism. The gene of cold tolerance at booting stage was further mapped on chromosome 5 between RM7452 and RM31 with genetic distances of 4.8 cM and 8.0 cM, respectively. This gene explained 10.50% of phenotypic variance and 5.10% of phenotypic variance of fully filled grains, and was tentatively designated as Ctb（t）.     

水稻抽穗扬花期耐热QTL(qHTH5)定位及其遗传效应分析

对元江普通野生稻(Oryza rufipogon Griff.)(简称元江普野)荷花塘3号为供体、籼稻(O.sativassp.indica)恢复系蜀恢527为轮回亲本构建的种间近等基因系群体进行数量性状位点(QTL)分析,在近等基因导入系YJ10-03-01中鉴定了一个抽穗扬花期耐热QTL。利用均匀分布在水稻12条染色体上的360个SSR标记检测近等基因系YJ10-03-01和籼稻恢复系蜀恢527,共获得9个多态性标记。单标记分析表明第5染色体短臂上的多态性标记与抽穗扬花期耐热性极显著相关。进一步在人工气候室模拟高温条件下处理YJ10-03-01与蜀恢527杂交得到F2分离群体(1027个单株)并进行SSR标记分析,以水稻结实率为耐热指标,利用复合区间作图方法在第5染色体短臂上检测到一个抽穗扬花期耐热性QTL,暂命名为qHTH5。该QTL在F2及F3世代分别解释8.6%和19.4%的表型变异。在F3世代,继续利用目标区间标记RM7320和RM7444之间的SSR标记鉴定纯合重组体,利用置换作图法将QTL定位在约304.2kb之内(RM592-RM17921)。     

Haplotyping of Rice Genotypes Using Simple Sequence Repeat Markers Associated with Salt Tolerance

Salt stress is a major problem in most of the rice growing areas in the world. A major QTL Saltol associated with salt tolerance at the seedling stage has been mapped on chromosome 1 in rice. This study aimed to characterize the haplotype diversity at Saltol and additional QTLs associated with salt tolerance. Salt tolerance at the seedling stage was assessed in 54 rice genotypes in the scale of 1 to 9 score at EC = 10 d Sm-1 under controlled environmental conditions. Seven new breeding lines including three KMR3/O. rufipogon introgression lines showed similar salt tolerant ability as FL478 and can be good sources of new genes/alleles for salt tolerance. Simple sequence repeat(SSR) marker RM289 showed only two alleles and RM8094 showed seven alleles. Polymorphic information content value varied from 0.55 for RM289 to 0.99 for RM8094 and RM493. Based on 14 SSR markers, the 54 lines were clearly separated into two major clusters. Fourteen haplotypes were identified based on Saltol linked markers with FL478 as the reference. Alleles of RM8094 and RM3412 can discriminate between the salt tolerant and susceptible genotypes clearly and hence can be useful in marker-assisted selection at the seedling stage. Other markers RM10720 on chromosome 1 and RM149 and RM264 on chromosome 8 can also distinguish tolerant and susceptible lines but with lesser stringency.     

Genetic diversity and association mapping for salinity tolerance in Bangladeshi rice landraces

Breeding for salinity tolerance using Bangladeshi rice landraces and understand genetic diversity has been limited by the complex and polygenic nature of salt tolerance in rice genotypes. A genetic diversity and association mapping analysis was conducted using 96 germplasm accessions with variable response to salt stress at the seedling stage. These included86 landraces and 10 indica varieties and lines including Nona Bokra, from southern Bangladesh. A total of 220 alleles were detected at 58 Simple Sequence Repeat(SSR) marker loci randomly distributed on all 12 rice chromosomes and 8 Sequence Tagged Site(STS) markers developed for genes SKC1, DST, and SalT. The average gene diversity was 0.5075 and polymorphism information content value was 0.4426, respectively. Cluster analysis revealed that 68 and 21 accessions were clustered into 2 distinct groups, possibly corresponding to indica and japonica groups, respectively and the remaining 7 landraces were classified as an admixed group. In addition to Wn11463, the STS marker for SKC1, RM22418 on Chr. 8 was significantly associated with salinity tolerance, at the location of a QTL detected in previous studies. Our findings of favorable alleles associated with salinity tolerance in Bangladeshi rice landraces, as well as the development of STS markers for salt tolerance genes, will be helpful in future efforts to breed salinity tolerance in rice.     

Identification of Rice Accessions Associated with K~+/Na~+ Ratio and Salt Tolerance Based on Physiological and Molecular Responses

The key for rice plant survival under Na Cl salt stress is maintaining a high K~+/Na~+ ratio in its cells. Selection for salt tolerance rice genotypes based on phenotypic performance alone will delay in progress in breeding. Use of molecular markers in tandem with physiological studies will help in better identification of salt tolerant rice accessions. Eight rice accessions along with the check Dongjin were screened using 1/2 Yoshida solution with 50 mmol/L NaCl at the seedling stage. The accessions IT001158, IT246674, IT260533 and IT291341 were classified as salt tolerant based on their K~+/Na~+ ratios. Seventeen SSR markers reported to be associated with K~+/Na~+ ratio were used to screen the accessions. Five SSR markers(RM8053, RM345, RM318, RM253 and RM7075) could differentiate accessions classified based on their K~+/Na~+ ratios. Banding pattern of the accessions was scored compared to the banding pattern of Dongjin. The study differentiated accessions based on their association of K~+/Na~+ ratio with molecular markers which are very reliable. These markers can play a significant role in screening large set of rice germplasms for salt tolerance and also help in identification of high-yielding varieties with better salt tolerance. The salt tolerant accessions can be taken forward into developing better varieties by conventional breeding and exploring genes for salt tolerance.     

Genetic Analysis and Gene Mapping of a Rice Tiller Angle Mutant tac2

Tiller angle, a very essential agronomic trait, is significant in rice breeding, especially in plant type breeding. A tiller angle controlling 2 (tac2) mutant was obtained from a restorer line Jinhui 10 by ethyl methane sulphonate mutagenesis. The tac2 mutant displayed normal phenotype at the seedling stage and the tiller angle significantly increased at the tillering stage. A preliminary physiological research indicated that the mutant was sensitive to GA. Thus, it is speculated that TAC2 and TAC1 might control the tiller angle in the same way. Genetic analysis showed that the mutant trait was controlled by a major recessive gene and was located on chromosome 9 using SSR markers. The genetic distances between TAC2 and its nearest markers RM3320 and RM201 were 19.2 cM and 16.7 cM, respectively.     

水稻耐热性的QTL定位及耐热性与光合速率的相关性

应用典型的籼粳交组合IR64×Azucena花药培养的DH群体及其已构建的分子连锁图谱,在田间及温室高温条件下对该DH群体的结实性状进行考查,采用QTLmapper 1.0软件检测控制结实率的加性和上位性效应的QTL。在第1、3、4、8和11等5条染色体上,共检测到6个具有加性效应的QTL,其中位于第1、3染色体的2个加性效应QTL来自父本Azucena的等位基因,它们是耐热的QTL,能分别提高结实率9.50和6.46个百分点,其贡献率分别为19.15%和2.86%;位于其余3条染色体的4个加性效应的QTL来自母本IR64的等位基因,它能提高结实率4.33~10.37个百分点,在第1、2、3、4、5、7、8、11等8条染色体之间还检测到8对加性×加性上位性效应,其贡献率为2.27%~8.13%。同时还对水稻分蘖盛期和抽穗期进行了光合速率的测定,发现抽穗期剑叶光合速率与耐热性呈显著的正相关。