普通野生稻耐冷性研究进展

普通野生稻是栽培稻的近缘种,含有许多特异基因,耐冷性突出,有利于耐冷性水稻品种的培育。综述了普通野生稻耐冷基因的定位与克隆、耐冷生理生化和分子机制及其在水稻耐冷性改良中的应用等研究进展,对今后普通野生稻耐冷性相关研究作了展望。     

利用染色体片段置换系定位水稻芽期耐冷性QTL

以籼稻品种9311为受体、粳稻品种日本晴为供体构建的95个染色体片段置换系为材料,在5℃低温条件下进行芽期耐冷性鉴定。结果表明,6个置换系低温处理后的成苗率与受体亲本9311有一定差异,其耐冷性略强于9311。利用代换作图法共鉴定出4个与芽期耐冷性相关的QTL,分别位于水稻第5和第7染色体上。其中qCTB-5-1、qCTB-5-2和qCTB-5-3分别被定位在第5染色体RM267与RM1237、RM2422与RM6054及RM3321与RM1054之间遗传距离分别为21.3cM、27.4cM和12.7cM的置换片段上;qCTB-7被定位在第7染色体RM11-RM2752区间遗传距离为6.8cM的置换片段上。     

利用重测序的染色体片段代换系群体定位水稻粒型QTL

 以一套用籼稻品种9311为受体、粳稻品种日本晴为供体构建的128个染色体片段代换系为材料,按随机区组实验设计种植,于成熟后考查代换系粒长、粒宽性状,利用多元回归分析方法,结合Bin图谱,鉴定出6个与粒长相关的QTL、2个与粒宽相关的QTL。其中,qGL3.1被定位在水稻第3染色体的5 792 954 bp区间内;qGL3.2被定位在第3染色体的917 878 bp区间内;qGL8.1被定位在第8染色体的889 543 bp区间内;qGL8.2被定位在第8染色体的208 614 bp区间内;qGL9.1被定位在第9染色体的1 149 685 bp区间内;qGL11.1被定位在第11染色体的3 184 760 bp区间内;qGW1.1被定位在第1染色体的200 070 bp区间内;qGW5.1被定位在第5染色体的704 905 bp区间内。上述QTL的准确定位,为进一步精细定位及克隆相应QTL和开展水稻粒型分子育种奠定了基础。     

不同温光条件下水稻抽穗期QTL的定位与分析

以广陆矮4号为受体,日本晴为供体的85个染色体单片段代换系群体为试验材料,通过单因素方差分析和Dunnett多重比较,测验单片段代换系与广陆矮4号之间抽穗期的差异,对代换片段上抽穗期相关的QTL进行了定位。以P≤0.001为阈值,在南京和海南不同温光条件下共定位到40个抽穗期相关的QTL。其中,21个QTL在2个环境中均被检测到;15个QTL只在南京环境中被检测到;4个QTL只在海南环境中被检测到。南京环境中定位到的36个抽穗期相关QTL,其加性效应值变化范围为2.8d~15.7d,加性效应百分率变化范围为3.8%~21.1%;海南环境中定位到的25个抽穗期相关QTL,加性效应值变化范围为1.8d~12.1d,加性效应百分率变化范围为1.7%~11.3%。这些QTL的定位,为进一步精细定位并克隆相应主效QTL和优异品种特定环境下的生育期改良奠定了基础。     

云南稻种冲腿的孕穗期耐冷性QTL分析

 利用352 个探针分析了日本品种农林20和云南品种冲腿之间的限制性片段长度多态性（RFLP），从中筛选出157个在农林20和冲腿间显示多态性的探针，其中55个被用于其杂交后代（农林20/冲腿）的70个F3系统的孕穗期耐冷性数量性状位点（QTL）分析。结果初步显示出一些与水稻孕穗期耐冷性有关的QTL，它们主要分布在第1、3、4、5、6、7、8、10和第12染色体上。可能在第3和第7染色体上具有对孕穗期耐冷性作用较大的QTL。     

水稻苗期耐冷性QTLs的定位

利用耐冷的籼稻资源743与冷敏的广亲和材料Dular杂交得到的86个重组自交系(RIL)为材料,构建了一张包含90个微卫星标记的水稻分子连锁图谱.以12℃低温处理下叶绿素含量、丙二醛含量的变化和4℃致死温度处理后的凋萎率为耐冷性指标,进行了苗期耐冷性数量性状位点(QTLs)的分析.以低温下叶绿素含量为指标,检测到分别位于染色体3,5,6(2个)和9上的5个QTIs;而在染色体2,3,9和12上检测到与低温下丙二醛含量有关的4个QTLs;用凋萎率为指标定位的5个QTLs则分别位于染色体1(2个),3,9和11上.这些QTLs控制的表型变异分别为3.07%～17.15%.位于染色体9上的RM105-RM257区间的QTL与低温下的叶绿素含量、丙二醛含量及植株凋萎率均有关,是控制苗期耐冷性的主效QTL.     

基于水稻单片段代换系的粒形QTL定位

 粒形是重要的农艺性状,既影响水稻产量也影响稻米品质,是典型的数量性状,易受遗传背景和环境因素的影响。染色体单片段代换系降低了遗传背景的干扰,是鉴定QTL的新型遗传材料。利用单片段代换系为材料,共鉴定出了22个与粒形相关的QTL,包括7个粒长QTL,6个粒宽QTL,5个谷粒长宽比QTL和4个粒厚QTL,分布于除第6、9、11和12染色体外的8条染色体上。这些结果为粒形QTL的克隆和分子育种奠定了基础。     

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。     

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。     

玉米株高qPH10的QTL定位及候选基因预测

利用掖478为轮回亲本、齐319为供体亲本构建的染色体片段代换系CL137为父本,与掖478杂交构建近等基因系F2分离群体,根据齐319、掖478重测序数据开发在双亲中具有多态性的Indel分子标记,在两个环境中对控制玉米株高的10号染色体QTL进行定位。结果表明,2017年的株高表型将QTL定位到标记mk8-bnlg1655之间,位于83.86~85.34 Mb(B73 RefGen_v3)的1.5 Mb区间,表型贡献率为7.57%;2018年株高表型将株高QTL定位到标记mk5-bnlg1655之间,位于82.76~85.34 Mb的2.5 Mb区间,表型贡献率为5.75%。同时检测发现,该QTL主要以加性效应为主,显性效应较小。通过对所定位的QTL重合区间内的基因进行功能注释,预测可能控制株高的候选基因,为后续精细定位第10号染色体株高QTL以及探索候选基因功能机制提供研究基础。     

利用重测序的水稻染色体片段代换系定位控制稻米淀粉黏滞性谱QTL

 稻米RVA谱是评价稻米蒸煮与食用品质的重要指标之一,发掘新的控制稻米RVA谱QTL对稻米品质改良具有重要意义。利用以粳稻品种日本晴为受体、籼稻品种9311为供体并经高通量重测序的染色体片段代换系群体为材料,在两年两点的环境下对该群体中控制RVA谱特征值的QTL进行了定位分析。通过单因素方差分析和Dunnett多重比较, 分析染色体片段代换系与受体亲本之间相关RVA谱特征值的差异,以两年两点都能检测到的显著差异位点作为稳定表达的QTL。共检测到10个稳定表达的QTL,包括控制峰值黏度的4个QTL qPKV2.1、qPKV5.1、qPKV7.1和qPKV8.1,控制热浆黏度的2个QTL qHPV5.1和qHPV7.1,控制冷胶黏度的2个QTL qCPV5.1和qCPV7.1及控制消减值的2个QTL qSBV2.1和qSBV7.1。在两年两点的检测中10个稳定表达QTL的贡献率介于-31.8%～53.2%,这10个QTL分布在第2、5、7和8染色体上,其中位于第2、5和7染色体上的位点存在一因多效性。     

Detection of QTL for Cold Tolerance at Bud Bursting Stage Using Chromosome Segment Substitution Lines in Rice (Oryza sativa)

The cold tolerance at the bud bursting stage (CTB) was evaluated at 5°C by using a set of 95 chromosome segment substitution lines (CSSLs) derived from an indica rice 9311 and a japonica rice Nipponbare with a genetic background of 9311. The result showed that six CSSLs had slightly stronger effect on CTB than 9311. Total four quantitative trait loci (QTLs) for CTB were preliminary mapped on chromosomes 5 and 7 by substitution mapping. qCTB-5-1, qCTB-5-2 and qCTB-5-3 were mapped in the region of RM267-RM1237, RM2422-RM6054 and RM3321-RM1054, which were 21.3 cM, 27.4 cM and 12.7 cM in genetic distance on rice chromosome 5, respectively. qCTB-7 was mapped in a 6.8-cM region of RM11-RM2752 on rice chromosome 7.     

Identification of QTLs for Cadmium Tolerance During Seedling Stage and Validation of qCDSL1 in Rice

Cadmium(Cd)is a non-essential toxic metal that is harmful to plants.To investigate the genetic mechanism of Cd tolerance in rice,quantitative trait loci(QTLs)associated with Cd tolerance at the seedling stage were analyzed using a recombinant inbred line(RIL)population derived from a cross between PA64s and 93-11.A total of 36 QTLs associated with shoot length,root length,shoot dry weight,root dry weight and total dry weight were detected in Hangzhou and Lingshui of China.Among them,15 QTLs were identified under the control condition and 15 QTLs were identified under the Cd stress condition,and 6 QTLs for Cd tolerant coefficient were detected on chromosomes 1,3,7 and 9.The qCDSL1.1 and qCDSL1.2 were identified in Hangzhou and Lingshui,respectively,and had overlapping intervals on chromosome 1.To further confirm the effects of qCDSL1.1 and qCDSL1.2,we developed a chromosome segment substitution line(CSSL),CSSLqCDSL1,in 93-11 background harboring qCDSL1.1/qCDSL1.2 from PA64s.Compared to 93-11,CSSLqCDSL1 had increased shoot length under the Cd stress condition.These results pave the way for further isolation of those genes controlling Cd tolerance in rice and marker-assistant selection of rice elite varieties with Cd tolerance.     

Mapping QTLs for Drought Tolerance at Seedling Stage in Rice Using Doubled Haploid Population

QTLs for drought tolerance at the rice seedling stage were analyzed using a doubled haploid (DH) population consisted of 251 lines from the cross between a japonica parent Maybelle and an indica parent Baiyeqiu. A genetic linkage map with 226 SSR marker loci was constructed. Single-locus analysis following composite interval mapping (CIM) detected a total of five QTLs located on five different chromosomes of rice. Four QTLs were also detected following two-locus analysis, resolving two pairs of epistatic QT...     

以广东高州普通野生稻为供体亲本的水稻单片段代换系构建

以栽培稻品种粤香占为受体亲本,利用杂交、回交和微卫星标记辅助选择相结合的方法,构建了以高州普通野生稻为供体亲本的水稻单片段代换系群体。该群体由20个编号的9个单片段代换系构成。这9个单片段代换系分别分布在水稻的第1、2、3、10和11染色体上,代换片段长度为8.1～23.8cM,总长度为152.7cM,平均长度为17.0cM,代换片段对水稻基因组的覆盖总长度为136.1cM,覆盖率为7.5%。构建以高州普通野生稻为供体亲本的水稻单片段代换系,为发掘和利用高州普通野生稻的有利基因提供了理想的试验材料,为进一步开展高州普通野生稻的遗传研究提供了一条新途径。     

QTL Analysis for Yield Traits Related to Low Nitrogen Tolerance Using Backcrossing Recombinant Inbred Lines Derived from Dongxiang Wild Rice (Oryza rufipogon Griff.)

【Objective】Dongxiang wild rice (Oryza rufipogon Griff.) has strong low nitrogen tolerance and is an important germplasm for low nitrogen tolerance improvement. Identification of genes responsible for low nitrogen tolerance in Dongxiang wild rice is of great importance to understand molecular mechanisms of low nitrogen tolerance and develop rice varieties with low nitrogen tolerance. 【Method】Quantitative trait loci (QTLs) for plant height and yield traits under low and normal nitrogen conditions was identified using backcrossing recombinant inbred lines (BC1F12) derived from an interspecific cross Xieqingzao B // Dongxiang wild rice/Xieqingzao B and its genetic linkage. 【Result】A total of 57 QTLs were detected in 33 regions on all chromosome, except chromosome 4 and 8. They explained individually 3.17%~63.40% phenotypic variation, and 32 QTLs of them had favorable alleles derived from Dongxiang wild rice. Nineteen QTLs were simultaneously detected under both nitrogen treatments, and 38 QTLs were only identified under single nitrogen treatment, suggesting various genetic mechanisms in rice growth and yield formation under low and normal nitrogen conditions. 【Conclusion】Fourteen QTL clusters, 43 QTLs included, scattered on seven chromosomes, indicating the common genetic-physiological mechanisms behind different traits, and the QTL pyramiding for low nitrogen tolerance can be achieved by molecular marker-assisted selection.     

Identification of QTLs for Plant Height and Its Components by Using Single Segment Substitution Lines in Rice （Oryza sativa）

QTLs for plant height and its components on the substituted segments of fifty-two single segment substitution lines （SSSLs） in rice were identified through t-test （P〈0.001） for comparison between each SSSL and recipient parent Huajingxian 74. On the 14 substituted segments, 24 QTLs were detected, 10 for plant height, 2 for panicle length, 4 for length of the first internode from the top, 5 for length of the second internode from the top and 3 for length of the third internode from the top, respectively. All these QTLs were distributed on nine rice chromosomes except chromosomes 5, 9 and 11. The additive effect ranged from -4.08 to 3.98 cm, and the additive effect percentages varied from -19.35% to 10.43%.     

水稻耐冷性遗传及基因定位研究概况与展望

综述了水稻低温发芽力,幼苗期、孕穗期和开花期耐冷性等遗传研究概况和近年来在水稻耐冷性数量性状基因定位(QTLs)研究方面所取得的进展,并展望了今后的水稻耐冷性研究方向。     

Identification of Rice QTLs for Important Agronomic Traits with Long-Kernel CSSL-Z741 and Three SSSLs

Rice kernel shape affects kernel quality (appearance) and yield (1000-kernel weight) and therefore is an important agronomic trait, but its inheritance is complicated. We identified a long-kernel rice chromosome segment substitution line (CSSL), Z741, derived from Nipponbare as a recipient and Xihui 18 as a donor parent. Z741 has six substitution segments distributed on rice chromosomes 3, 6, 7, 8 and 12 with an average replacement length of 5.82 Mb. Analysis of a secondary F₂ population from a cross between Nipponbare and Z741 identified 20 QTLs for important agronomic traits. The kernel length of Z741 is controlled by a major QTL (qKL3) and a minor QTL (qKL7). Candidate gene prediction and sequencing indicated that qKL3 may be an allele of OsPPKL1, which encodes a protein phosphatase implicated in brassinosteroid signaling, and qKL7 is an unreported QTL. Finally, we validated eight QTLs (qKL3, qKL7, qRLW3-1, qRLW7, qPH3-1, qKWT3, qKWT7 and qNPB6) using three selected single- segment substitution lines (SSSLs), S1, S2 and S3. Also, we detected five QTLs (qKL6, qKW3, qKW7, qKW6 and qRLW6) in S1, S2 and S3, which were not found in the Nipponbare/Z741 F₂ population. However, qNPB3, qNPB7 and qPL3 QTLs were not validated by the three SSSLs in 2019, suggesting that minor QTLs are susceptible to environmental factors. These results lay the foundation for studying the biodiversity of kernal length and molecular breeding of different kernel types.