水稻株高QTL及其与产量性状和抽穗期关系的研究进展

株高是一个与水稻品种丰产潜力密切相关的重要性状。主效半矮秆基因背景下的水稻株高变异,一般表现为受多基因控制的数量性状。最近的研究表明,已定位的株高QTL分布于水稻的所有12条染色体,其中4个QTL已克隆。克隆研究和QTL初定位结果表明,株高QTL往往存在对产量性状和（或）抽穗期的多效作用,可利用于提高水稻产量潜力。     

Comparative proteomics analysis of maize (Zea mays) leaves infected by small brown planthopper (Laodelphax striatellus)

Maize rough dwarf disease (MRDD) is a viral disease caused by brown planthopper infestation, and leads to great yield loss, especially in China. Comparative proteomics was performed using maize inbred line Zheng 58 and LN 287. MRDD pathogen was detected as rice black-streaked dwarf virus (RBSDV) by quantitative real time PCR (qRT-PCR) in Shandong Province, China. The modified trichloroacetic acid (TCA)/acetone method was used for soluble protein extraction from leaves. Two-dimensional electrophoresis (2-DE) analysis was performed on 24-cm long, pH 4–7 linear immobilized pH gradient (IPG) strips, and gels were stained with silver and coomassie brilliant blue. We identified 944 proteins expressed in RBSDV infected maize leaves by proteomics approaches. Among these, 44 protein spots that revealed a 1.5-fold difference in intensity were identified by mass spectrometry between mock-inoculated and RBSDV infected samples. Among these, 17 and 26 spots were up-regulated, and 27 and 18 spots were down-regulated in the virus infected samples of Zheng 58 and LN 287, respectively. Differential protein spots were analyzed by mass spectrometry identification, which could be divided into six categories. Furthermore, the expression of stress-related proteins was detected and confirmed by qRT-PCR. This study lays the foundation for further investigations, enabling the enhancement of MRDD resistance in maize.     

大豆脱氢抗坏血酸还原酶基因的电子克隆及进化分析

利用生物信息学和实验验证的技术路线,以拟南芥脱氢抗坏血酸还原酶基因cDNA序列为信息探针,对大豆EST数据库进行同源搜索,经同源性比对和序列组装,获得全长为955bp的大豆脱氢抗坏血酸还原酶基因的cDNA序列(GenBank登录号为DQ006810),经RT-PCR扩增,分子克隆和序列分析验证,结果表明与电子克隆完全一致.该cDNA序列具有完整的开放阅读框架(ORF,36bp-815bp),推测编码蛋白为259个氨基酸.通过与几种植物脱氢抗坏血酸还原酶蛋白序列的比较,发现该基因具有较高的保守性.结果表明根据物种问同源基因序列,对跨物种问EST数据库进行同源检索筛选,拼接,是新基因克隆的一条有效途径.     

富含多糖甜玉米幼穗RNA提取及高温胁迫基因表达分析

以改良Trizol试剂法从富含多糖的甜玉米幼穗中提取高质量、完整的总RNA,利用实时荧光定量PCR对高温胁迫下粤甜13雌穗发育的8个差异表达基因进行分析。结果表明,8个基因分为上调表达和下调表达,实时荧光定量PCR和测序法基因表达谱检测的基因上调或下调表达的趋势一致,上调表达基因ZM2G058057和下调表达基因ZM2G059964、ZM2G044670相对表达量较高,可能在高温胁迫影响甜玉米幼雌穗发育过程中起更重要的作用。     

Genetic studies for breeding of rice cultivars with superior grain appearance and lodging resistance from the rice cultivar ‘Emi-no-kizuna’

ABSTRACT

We conducted a quantitative trait locus (QTL) analysis of grain appearance (GA) and agronomic traits of rice, using 128 recombinant inbred lines derived from ‘Emi-no-kizuna’ and ‘Tomohonami’. We detected two promising QTLs associated with GA: qGA4 on chromosome 4 and qGA8 on chromosome 8. qGA4 contributed highly to the greater percentage of perfect grains of the Emi-no-kizuna genotype. In the same region, we detected other QTLs associated with panicle number and spikelet number per panicle. In near-isogenic lines (NILs) in which Emi-no-kizuna alleles were introgressed in the genomic region of only the semi-dwarf 1 (sd1) locus (NIL_1) and both the sd1 locus and qGA8 (NIL_2), respectively, the percentage of perfect grains was significantly higher and the percentages of milky white, basal white, and white back grains were significantly lower than in Tomohonami; and the percentages of milky white and white back grains of NIL_2 were significantly lower than those of NIL_1. These results suggest that introgression in the sd1 region could improve GA, and that the addition of qGA8 could further improve GA. The culm lengths of the NILs were significantly shorter than that of Tomohonami, indicating improved lodging resistance. Grain weight of NIL_2 was significantly smaller than that of NIL_1, suggesting that the effect of qGA8 could be pleiotropic, or the gene that underlies qGA8 could be linked with genes associated with grain weight.

Abbreviations: ANOVA: analysis of variance; AT20: mean air temperature in the 20 days after heading; BW: basal white grain; CL: culm length; DAH: days after heading; GA: grain appearance; GW: 1000-grain weight; LOD: logarithm of odds; MW: milky white grain; NIL: near-isogenic line; PG: perfect grain; PL: panicle length; PN: panicle number; PTSN: putative total spikelet number; PVE: percentage of phenotypic variation explained; QTL: quantitative trait locus; RIL: recombinant inbred line; SN: spikelet number per panicle; SNP: single nucleotide polymorphism; WB: white back grain     

小麦籽粒淀粉合成酶基因表达与酶活性特征的研究

为研究小麦籽粒淀粉合成酶基因表达与酶活性的特征,以普通小麦品种秦麦11(粒重36.942 mg/粒,淀粉含量61.02%)和中优9507(粒重50.636 mg/粒,淀粉含量68.36%)为材料,采用实时荧光定量RT-PCR方法,对灌浆期籽粒淀粉合成酶相关基因的表达进行了研究,并对其表达量与相应酶活性的相关性做了分析.结果表明,腺苷二磷酸葡萄糖焦磷酸化酶基因(AGPase)、束缚态淀粉合成酶基因(GBSSI)、可溶性淀粉合成酶基因(SSS)、淀粉分支酶基因(SBE)表达均呈单峰曲线变化,在花后3 d 内检测不到其表达,花后4～6 d 这些基因开始表达.AGPase和SSS在花后12 d左右表达量达到高峰,GBSSI和SBE在花后15 d左右的表达量最大.自花后18 d开始,各基因的表达量均显著下降.中优9507在表达高峰期(即花后第12、15、18 d)的酶基因相对表达量均显著高于秦麦11,且差异均达显著水平.整个灌浆期间中优9507的四种淀粉合成酶活性高于秦麦11,尤其在灌浆中期差异更显著.相关分析表明,AGPase、SSS、SBE基因在花后15 d的相对表达量与相应酶活性间呈极显著正相关,而GBSS基因的相对表达量与GBSS酶活性间相关不显著,暗示AGPase、SSS、SBE基因在籽粒淀粉合成过程中属于转录水平调控,而GBSSI基因属于转录后调控.     

高效液相色谱法测定香蕉防霜剂中三氯卡班的含量

采用高效液相色谱法,色谱条件为：Symmetry十八烷基键合硅胶柱,流动相：乙腈∶水=7∶3（V∶V）,检测波长：265 nm,流速：1 mL.min-1。建立了测定香蕉防霜剂中三氯卡班含量的方法,线性方程为：Y=5.02×10-4X＋1.27×10-3（Y为三氯卡班含量,X为色谱峰面积,R=0.9997）,线性范围为0.8-15 mg.L-1,检测限0.2 mg.L-1,方法回收率为99.08%-99.32%（RSD=1.08%-2.03%,n=5）。可用于香蕉防霜剂和相关产品中三氯卡班定量测定。     

番木瓜果实中芥子酶基因组织差异表达和酶活性研究

检测了番木瓜果实3个发育时期(Ⅰ、Ⅱ、Ⅲ)的果皮(外果皮)、果肉(内果皮)及种子中芥子酶的活性,并通过RT-PCR分析CpTGG1、CpTGG2和 CpTGG3三个番木瓜芥子酶基因在番木瓜果实各组织的差异性表达情况。结果表明,番木瓜果皮及种子中均有明显的芥子酶活性,但在果肉中无法检测出芥子酶活性;成熟后(Ⅲ期)番木瓜果皮的芥子酶活性相比未成熟前提高了6.6倍,而不同发育时期种子中的芥子酶并不表现出显著差异性。RT-PCR分析结果表明,番木瓜芥子酶基因在番木瓜果实中的表达存在显著时空差异性。其中,CpTGG1在番木瓜果皮及种子中均有表达,且表达量无明显差异,在果肉中完全无表达;CpTGG2在番木瓜果实的各个组织中均无表达;CpTGG3在果皮及种子中也均有表达,成熟果皮(Ⅲ期)和Ⅰ期种子中表达量比较强,并随着果实的发育成熟而呈现梯度变化。     

水稻种子低氧发芽力的QTL定位和上位性分析

利用35份水稻品种资源,评价了在不同水深、温度等低氧条件下的种子萌发情况,表明水温30℃、水深0.2 m下黑暗萌发5 d为最佳鉴定方法,并以此条件下水稻种子萌发的芽长为鉴定指标,评价了359份水稻品种低氧发芽力地区间、籼粳间的差异。进一步利用81个Kinmaze/DV85 重组自交系群体进行了水稻低氧发芽力数量性状位点分析, 在第1、2、5、7染色体上检测到5 个低氧发芽力QTL,其中第5染色体上存在2个QTL,各QTL的贡献率为10.5%～19.6%。同时进行上位性分析,检测到3对互作位点,分别位于第2、3、5、11染色体上,其中位于第3染色体上标记C563-X182之间的位点与第5染色体上标记R830-X208之间的位点的互作贡献率高达48.78%。     

关于植物群丛划分的探讨

在传统的植物群落分类系统中,群丛是植物群落分类的基本单位.从群丛分类的必要性出发,综述了传统植物群落分类系统中对群丛的定义及其划分方法,即在群丛的划分中主要依据群落中不同层片的优势种或特征种;但是在利用传统植物群落分类方法划分群丛时也存在一些不确定性因素,主要表现在确定群丛的特征种(组)时需要人为确定;同时,论述了当前植物群落数量分类的研究现状,分析了利用双向指示种分析法(TWINSPAN)、主成分分析(PCA)等数量分类方法划分群丛时存在的一些问题,主要表现在数量分类结果与传统分类单位的对应关系不能达到协调一致,无法判断是否划分到了群丛的水平.最后提出了群丛划分方法的展望:数量方法是基础,特征种(组)是及其数量特征是关键.