控制水稻金黄色颖壳与节间基因OsCAD2的图位克隆

【目的】水稻色素不仅对其自身生长发育有重要生理作用,而且在水稻育种、农副产品改良等方面运用比较广泛。对水稻色素相关基因进行表型分析和基因定位,为进一步研究色素代谢途径及调控机理奠定基础。【方法】利用EMS诱变粳稻长粒粳(CLJ),在突变体库内筛选到一个金黄色颖壳与节间突变体gh881;在成熟期,测定野生型与gh881的主要农艺性状;将gh881与野生型及中恢8015杂交,观察BC1F1及F1植株表型,并对BC1F2及F2表型分离进行卡方检验,对gh881进行遗传分析;利用F2群体和图位克隆的方法对gh881突变基因进行定位;采用q PCR检测颖壳颜色相关基因在gh881与野生型不同发育时期的幼穗、节间以及剑叶叶鞘的相对表达量。【结果】与野生型相比,突变体的颖壳与节间均呈金黄色;除单株有效穗数外,gh881突变体的株高、每穗总粒数及实粒数、结实率和千粒重等性状均极显著降低。遗传分析和基因定位结果表明,gh881的突变表型受1对隐性核基因控制,位于第2染色体短臂,并最终将该基因精细定位于标记FH-13和RH-25之间,物理距离约33.2 kb,该区域中包含4个开放阅读框(ORFs)。【结论】序列分析结果表明,发现其中一个编码肉桂醇脱氢酶(CAD)的基因OsC AD2(Os02g0187800)的3 563 bp处发生了1个单碱基突变(G转换为A),导致该基因编码区的第297位氨基酸由甘氨酸突变为天冬氨酸,由此认为该突变体为OsC AD2基因单碱基突变的新等位基因。q RT-PCR结果表明,突变体的节间中OsCAD2相对表达量极显著下调,而在剑叶叶鞘及穗部则基本都是极显著增加,其他相关基因也发生显著变化,证实OsCAD2是木质素代谢中的重要基因,且可能与其他相关基因存在反馈调节。     

水稻叶片内卷突变体rl(t)的鉴定与基因定位

【目的】叶片是水稻理想株型的重要内容,叶片适度卷曲可以提高光合效率。对卷叶相关基因进行遗传分析和初步定位,为下一步的基因克隆与功能分析提供研究基础。【方法】利用EMS诱变雄性不育保持系宜香1B获得一份稳定遗传的叶片向内卷曲突变体,暂命名为rl(t)。在成熟期,测定野生型和rl(t)的主要农艺性状;在分蘖期,取野生型和rl(t)叶片用FAA固定液固定进行石蜡切片,同时,用野生型和rl(t)剑叶测定叶绿素含量;在抽穗期,利用Li-6400便携式光合仪测定10株抽穗期的野生型和rl(t)的光合参数;将rl(t)与野生型及日本晴杂交,观察F_1植株表型,对F_2表型分离进行χ~2测验,对突变体进行遗传分析。以rl(t)/日本晴的F_2群体为材料,利用BSA法进行定位。【结果】与野生型相比,突变体叶片向内卷曲明显,叶片更加直立,叶色变深,其他主要农艺性状均有不同程度降低。光合特性分析表明,突变体比野生型具有更高的光合色素含量,但光合效率没有明显差异。叶片组织切片观察表明,突变体中泡状细胞变小可能是导致叶片卷曲的主要原因。遗传分析表明,该突变体受一对隐性核基因控制,利用突变体与日本晴的F_2群体进行基因定位,最终将该基因定位在第7染色体长臂InDel标记Ind3和Ind4间610 kb的物理区间。【结论】rl(t)叶片内卷是由于近轴面泡状细胞面积减小。RL(t)定位区间内未见卷叶相关基因报道,推测RL(t)可能是一对新基因。     

利用90K基因芯片进行小麦株高QTL分析

为给小麦株高标记辅助选择提供可供选择的分子标记,并进一步对株高QTL进行精细定位及相关基因克隆,以小麦骨干亲本周8425B和小偃81衍生的包含102个家系的RIL群体(F_8)为材料,利用90K芯片标记构建高密度遗传图谱,在3个环境下对株高进行QTL检测。结果表明,所构建的图谱含有9 290个SNP标记,覆盖了小麦21条染色体的63个连锁群,图谱总长3 894.64cM,平均标记密度为0.42cM。共检测到9个控制株高的QTL,分布于1B、4A、4D、6B、7A、7B和7D染色体上,变异解释率为2.23%~16.25%。QPh.nafu.4D、QPh.nafu.4A、QPh.nafu.1B-2与前人定位到的位置相同或相近。QPh.nafu.7A具有较大的LOD值(8.17)和变异解释率(14.69%),为主效QTL。QPh.nafu.6B、QPh.nafu.7B-1、QPh.nafu.7B-2均能在多个环境下使用多种QTL检测方法定位到,可能为新的较稳定的控制株高的QTL。     

小麦白斑突变体I30的特征特性及遗传分析

类病斑突变体是研究植物程序性死亡和抗病性的理想材料。为了丰富小麦斑点突变体的研究,对叠氮化钠诱变小麦品种陕农33产生的稳定遗传的白斑突变体I30进行了特征特性研究和遗传分析。结果表明,突变体I30从三叶期开始表现白色块斑和长条纹。锥虫蓝染色和DAB染色显示,I30斑点处出现细胞死亡和H_2O_2积累现象。透射电子显微镜观察表明,I30的叶绿体形状发生改变,数目减少,基粒垛叠高度无序,部分甚至降解。农艺性状调查结果表明,I30的株高、单株有效穗数、穗粒数、穗长和结实率与野生型间无显著差异,但千粒重、穗粒重、单株产量、旗叶长度和宽度显著低于野生型。遗传分析表明,I30由1对隐性核基因控制。利用BSA+660K基因芯片技术,将该基因定位于小麦6D染色体上,位于SSR分子标记Xcfd190和6DS-5之间,遗传距离分别为6.4cM和9.1cM。     

小麦雄性不育系AL18A育性恢复基因的QTL定位

为加快AL型杂交小麦的发展,以不育系AL18A、恢复系99AR144-1及二者杂交F2代群体为材料,选用SSR标记和分离群体分组分析法进行育性恢复基因的QTL定位。结果表明,育性恢复由主效和微效基因共同控制,采用复合区间作图法分析,在1B染色体上检测到了1个主效恢复基因QTLqRf-1B-1,在5AL染色体上检测到了1个微效QTLqRf-5A-1。qRf-1B-1位于SSR标记Xbarc8与Xgwm413之间,与两标记的遗传距离分别为0.85cM和2.00cM,LOD值为14.06,加性效应为18.87,可解释22.43%的表型变异;qRf-5A-1位于SSR标记Xgwm595与Xgwm410之间,与两标记的遗传距离分别为10.00cM和0.10cM,LOD值为3.18,加性效应为12.32,可解释5.44%的表型变异。     

小麦旗叶大小及穗部相关性状的QTL定位

为了发掘更多控制小麦旗叶大小及穗部相关性状的QTL,以兰考906和小偃81创制的133个F6~F7重组自交系为试验材料,在6个环境下利用SSR标记对旗叶大小及穗部相关性状进行QTL定位。结果表明,有202对SSR标记被用于构建遗传连锁图谱,图谱覆盖小麦21条染色体,全长1 678.93cM,标记间平均距离8.30cM。采用完备区间作图法共检测到30个QTL,分布在1B、2A、3D、4A、4B、4D、5D、6A、6B、6D和7D染色体上。其中,旗叶宽QTL有7个,穗长QTL有9个,小穗数QTL有5个,穗粒数QTL有5个,小穗着生密度QTL有4个,不同环境下单个QTL可解释的表型变异率为4.94%~23.14%,有14个QTL的表型贡献率大于10%,有8个QTL可在2个或2个以上环境中被检测到。其中,Qflw-4A在3个环境中被检测到,贡献率为10.13%~20.77%,是控制旗叶宽的稳定主效QTL;Qsl-4D.2在4个环境中被检测到,贡献率为12.58%~23.14%,是控制穗长的稳定主效QTL;Qker-5D在2个环境中被检测到,贡献率为11.44%~14.32%,是控制穗粒数的稳定主效QTL。这3个稳定主效QTL可作为改良叶宽和增加穗粒数的功能QTL作进一步研究。     

Investigation and genetic mapping of a Glomerella leaf spot resistance locus in apple

Apple Glomerella leaf spot (GLS) is a severe fungal disease that damages apple leaves during the summer in China. Breeding new apple varieties that are resistant to the disease is considered the best way of controlling GLS. Fine mapping and tightly linked marker are critically essential for the preselection of resistant seedlings. In this study, a population of 207 F₁ individuals derived from a cross between ‘Golden Delicious’ and ‘Fuji’ was used to construct a fine simple sequence repeat (SSR)‐based genetic linkage map. The position of R_gls, a locus responsible for resistance to GLS, was identified on apple linkage group (LG) 15 using SSR markers CH05g05 and CH01d08, which was adapted from a published set of 300 SSR markers that were developed using the bulked segregant analysis (BSA) method. These two SSR markers flanked the gene, and its recombination rate was 8.7% and 23.2%, respectively. A total of 276 newly developed SSR markers around the target region and designed from the genome apple assembly contig of LG15 were screened. Only nine of these were determined to be linked to the R_gls locus. Thus, a total of 11 SSR markers were in linkage with R_gls, and mapped at distances ranging from 0.5 to 33.8 cM. The closest marker to the R_gls locus was S0405127, which showed a genetic distance of approximately 0.5 cM. The first mapping of the gene R_gls was constructed, and the locations of the 11 effective primers in the ‘Golden Delicious’ apple genome sequence were anchored. This result facilitates better understanding of the molecular mechanisms underlying the trait of resistance to GLS and could be used in improving the breeding efficiency of GLS‐resistant apple varieties.     

Mapping and validation of the quantitative trait loci for leaf stay‐green‐associated parameters in maize

Functional stay‐green is generally regarded as a desirable trait of varieties in major crops including maize. In this study, we used an F_3:4 recombinant inbred line population with 165 lines from a cross between a stay‐green inbred line (Zheng58) and a model inbred line (B73) using 211 polymorphic simple sequence repeat markers to map quantitative trait loci for three stay‐green‐associated parameters, chlorophyll content, photosystem II photochemical efficiency and stay‐green area, at maturity stage, detected a total of 23 quantitative trait loci (QTL) on nine chromosomes. Single QTL explained 3.7–13.5% of the phenotypic variance. Additionally, we validated some important stay‐green QTL using a heterogeneous inbred family approach and found that the stay‐green‐associated parameters were significantly correlated with the plant yield. This study may contribute to a better insight into the regulatory mechanism behind leaf stay‐green in maize and a novel development of elite maize varieties with delayed leaf senescence through molecular marker‐assisted selection.     

Soybean plant height QTL mapping and meta‐analysis for mining candidate genes

Plant height is closely related to seed yield of soybean. The goal of this study was to identify important loci affecting soybean plant height using meta‐analysis based on a reference physical map. Plant height related to QTLs was mapped across eight years with a RIL population by WinQTLCart v2.5. 182 QTLs related to plant height of soybean from database and our research were collected, and each QTL was projected onto the soybean physical map by software BioMercator v2.1. The confidence interval of meta‐QTL ranged from 0.09 to 5.07 Mb, and the mean phenotypic variance ranged from 4.9% to 73.0%. Furthermore, 4,259 candidate genes were located in these consensus QTLs, and 40 of them were involved in the plant growth and stem elongation and annotated as plant hormone signal transduction (pathway ID ko04075) in KEGG pathway. These results would lay a foundation for fine mapping of QTLs/genes related to plant height and marker‐assisted selection for breeding in soybean.