利用C_0t-1 DNA对高秆野生稻和宽叶野生稻基因组的比较分析

为了揭示同为CCDD基因组类型的高秆野生稻和宽叶野生稻的起源和进化关系,用高秆野生稻C0t-1DNA作为探针,对其自身体细胞染色体与宽叶野生稻体细胞染色体进行荧光原位杂交并对其核型进行同源性聚类和比较分析。杂交结果显示宽叶野生稻和高秆野生稻同源性非常高,说明它们的亲缘关系十分接近,但区别也非常明显,两者的杂交信号存在显著不同。C0t-1DNA具有很强的种特异性和依赖基因组型的特异性,利用它进行荧光原位杂交能够更有效地研究物种基因组间的关系,为物种的划分和鉴定提供依据。另外,通过对高秆野生稻和宽叶野生稻两个异源四倍体基因组的中高度重复序列的比较分析,探讨了稻属异源四倍体可能的起源与进化机制。     

栽培稻与紧穗野生稻间整倍体后代的RFLP分析

 用164个RFLP探针对67个栽培稻02428(2n=24,AA)与两份紧穗野生稻材料(2n=24,CC)间的整倍体(2n=24)后代进行分析,证实其中60个为易位系。各易位系分别带有1～6个紧穗野生稻染色体片段,很多易位系具有相同的易位发生区域。易位的紧穗野生稻片段都很小,主要分布在远离着丝点的区域。还就整倍体后代中出现新杂交带型的可能原因进行了讨论。     

水稻叶片衰老基因LPS1的克隆与功能研究

【目的】早衰突变体是研究早衰机制的良好载体,对于探究早衰的遗传机理与作用机制及提高水稻的产量和品质具有重要作用。【方法】本研究利用EMS诱变获得了一个早衰突变体lps1,并对该突变体及其野生型进行表型观察、细胞学及组织化学分析、生理生化分析、遗传分析、基因定位和激素处理。【结果】lps1的叶片从3叶期开始发黄,成熟期株高、有效分蘖数、结实率、千粒重等极显著降低。电镜观察发现lps1叶表面光滑,硅质化突起和叶绿体数目减少、片层结构紊乱。生理生化分析表明lps1中有大量的活性氧积累,同时伴有蛋白质的降解、细胞膜的损伤以及大规模的细胞死亡。遗传分析表明该早衰表型受单隐性核基因控制,并且其在第5染色体上编码了一个泛素结合酶。亚细胞定位结果证明LPS1蛋白在细胞质与核中均有表达。外源激素处理发现,lps1对外源激素的处理更为敏感,且LPS1突变促进了ABA合成相关基因的表达。【结论】LPS1 突变使水稻ABA合成信号途径异常,进而引发H₂O₂等一系列与衰老相关生理指标的异常变动,导致lps1过早衰老,最终造成水稻产量严重降低。     

玉米C4型PEPC蛋白的生物信息学分析

利用生物信息学分析软件对GeneBank上注册的玉米C4型PEPC蛋白质序列进行氨基酸组成成分、功能域、二级结构、疏水性、亚细胞定位、导肽及进化树分析和预测.结果表明:玉米PEPC蛋白是等电点5.77的亲水性不稳定蛋白,包含一个CYTH保守结构域,定位于细胞质中,具有中央中间代谢功能,属于连接酶类.α螺旋和不规则卷曲是其蛋白质二级结构的主要结构元件,β折叠和伸展链散布其中;玉米PEPC与高粱、水稻等植物的PEPC亲缘关系较远,不属于同一分支.     

中、韩两国主要茶树品种基因组DNA多态性比较研究

利用RAPD技术结合类平均法聚类分析研究了中国、韩国和日本茶树品种资源共 4 6个样品的基因组DNA多态性。结果表明 ,19个有效随机引物对 4 6个样品共扩增出 2 0 0条RAPD带 ,平均每个引物 10 5条带 ,每个品种 4 4条带。在得到的 2 0 0条谱带中 ,多态性带达到 84 3 %。中国和韩国茶树品种DNA多态性分别为 86 2 %和 78 2 %。 4 6个品种之间的遗传距离为 0 2 79～ 0 65 4。聚类分析结果表明 ,4 6个供试品种可分成 4个类群 ;韩国茶树品种和日本的薮     

稻属和假稻属植物外稃表面乳突结构的研究

 用扫描电镜对稻属植物21个种和假稻属植物2个种的外稃表面乳突作了观察。根据乳突的形态特征，将乳突分为6类，即双峰乳突、多向双峰乳突、瘤峰乳突、四峰乳突、凹痕乳突和假稻属的嵌硅双峰乳突。根据双峰距、垭深、距/深比和峰角度等性状，将双峰乳突分为锐型和钝型的两种类型。对乳突特征与稻种分类的关系以及稃毛的变态问题进行了讨论。     

稻属AA染色体组8个种间SSR多样性与亲缘关系

选用平均分布于水稻基因组的30对SSR 引物,对AA染色体组8个野生稻种共42份材料的遗传多样性及遗传关系进行了研究。结果显示,本试验选取的30个SSR标记均具有多态性,多态性位点百分率为100%。30个多态性位点共扩增出的等位基因数为224, 每个位点可扩增出3～10个等位基因,平均7.47个;等位基因有效数(Ae)变幅为1.25～891,平均5.45。多样性指数中,Shannon多样性指数(I)为0.454～2.386,平均1.826;而Nei基因多样性指数变幅为0.199～0.888,平均0.774。系统聚类和带型分析结果表明,亚洲栽培稻(Oryza sativa) 与普通野生稻(O. rufipogon) 的亲缘关系最近,非洲栽培稻(O. glaberrima) 则与巴蒂野生稻(O. barthii) 关系最为密切,杂草稻(O. spontanea) 与普通野生稻 (O. rufipogon)、亚洲栽培稻(O. sativa) 之间有较近的亲缘关系,而展颖野生稻(O. glumaepatula)、长雄蕊野生稻(O. longistaminata)与AA组其他稻种之间的亲缘关系较远。     

Pyranodipyran Derivatives with Tyrosyl DNA Phosphodiesterase 1 Inhibitory Activities and Fluorescent Properties from Aspergillus sp. EGF 15-0-3

Four new benzodipyran racemates, namely (±)-aspergiletals A–D (3–6), representing a rare pyrano[4,3-h]chromene scaffold were isolated together with eurotiumide G (1) and eurotiumide F (2) from the soft-coral-derived fungus Aspergillus sp. EGF 15-0-3. All the corresponding optically pure enantiomers were successfully separated by a chiral HPLC column. The structures and configurations of all the compounds were elucidated based on the combination of NMR and HRESIMS data, chiral separation, single-crystal X-ray diffraction, quantum chemical ¹³C NMR, and electronic circular dichroism calculations. Meanwhile, the structure of eurotiumide G was also revised. The TDP1 inhibitor activities and photophysical properties of the obtained compounds were evaluated. In the TDP1 inhibition assay, as a result of synergy between (+)-6 and (−)-6, (±)-6 displayed strong inhibitory activity to TDP1 with IC₅₀ values of 6.50 ± 0.73 μM. All compounds had a large Stokes shift and could be utilized for elucidating the mode of bioactivities by fluorescence imaging.     

海南普通野生稻自然居群间遗传多样性的微卫星分析

采用48个SSR标记分析了海南11个普通野生稻自然居群的遗传多样性。结果显示海南普通野生稻自然居群具有较丰富的遗传变异（Na=8.3,He=0.716）。居群内多数等位基因（77.6％）频率较低,其中,70个等位基因仅出现在1个居群中。62.9％的等位基因显著偏离哈迪 温伯格平衡,多数居群及等位基因实际杂合度大于期望杂合度,表明居群内自交率低、杂合体过剩。居群间遗传变异差异明显,在11个自然居群中,崖城居群遗传多样性最低（Na=1.7,He=0348）,和庆、椰林B两居群遗传变异最为丰富（和庆：Na=4.0,He=0.577;椰林B： Na=4.0,He=0.531）。Mantel测验表明,居群间Nei遗传距离与地理距离呈极显著相关（r=0.386,P=0004）。东、西海岸居群间遗传分化显著但较小（Fst=0.048,P=0024）,而居群间遗传分化则较大（Fst=0.335,P < 0.001）。11个自然居群间基因流非常有限（Nm=0.404）,表明居群间隔离程度较大。基于居群等位基因数目、基因多样性指数以及基因频率特点,认为海南11个自然居群中和庆和椰林B居群应是优先保护对象。     

一个水稻穗特异表达锌指蛋白基因的克隆与结构分析

利用生物信息学和RT PCR方法从水稻中克隆鉴定了一个新的具有TFⅢA型锌指结构的锌指蛋白基因,其开放阅读框为1092 bp,编码363个氨基酸残基。组织表达谱结果表明,该基因只在水稻幼穗组织中表达,在成株期的根、叶、茎以及花药中不表达,将其命名为OsZPT3 1。序列分析表明OsZPT3 1具有3个C2H2型锌指结构,在氨基酸的C端没有典型的转录抑制区域DLN box,但LXLXL的结构仍然表明OsZPT3 1可能是一个转录抑制因子。对OsZPT3 1启动子区域进行预测,结果发现3个MADS box转录因子识别位点,推测OsZPT3 1可能在MADS box转录因子的调节下,通过抑制下游基因的表达在水稻穗部器官的生长发育过程中发挥重要的调控作用。     

Comparative Embryological Studies on Infertility of Interspecific Hybridizations Between Oryza sativa with Different Ploidy Levels and O. officinalis

As maternal parents, diploid (L202-2x) and autotetraploid (L202-4x) of Oryza sativa cv. L202 were crossed with O. officinalis. Embryo development and fertilization in these two crosses were comparatively studied. There were no mature hybrid seeds obtained because all the hybridized spikelets died 30 days after pollination. The main reasons for no seed set were abnormal fertilization and development of the embryos and endosperms in the interspecific hybrids. There were double- fertilization, egg cell single-fertilization and non-fertilization in these crosses. Although 59.45% and 54.87% of hybrid embryos produced in the crosses of L202-2x/O. officinalis and L202-4x/O. officinalis, respectively, hybrid embryos ceased to develop or degenerated and plenty of free endosperm nuclei were in disaggregating state without developing cellular endosperms three days after pollination. Besides, some embryological differences in these two crosses were found, that is, the rate of double-fertilization and total rate of double- and single-fertilization in L202-2x/O. officinalis were higher than those in L202-4x/O. officinalis. The embryo and endosperm of hybrids developed more slowly, and embryos and free endosperm nuclei were more severely degenerated in L202-4x/O. officinalis than in L202-2x/O. officinalis. Five days after pollination, a few of embryos in L202-2x/O. officinalis developed into pear-shaped ones, however, embryos in L202-4x/O. officinalis were all degenerated. Therefore, it is more difficult to obtain interspecific hybrids by wide crosses between autotetraploid of O. sativa and O. officinalis.     

S1选择法对玉米群体产量性状改良效果的比较分析

以产量性状及农艺性状作为研究对象,比较分析前人利用S1选择法实施群体改良获得的遗传增益、遗传变异及对玉米群体结构的影响。比较发现,S1选择法能够有效提高玉米群体产量和一般配合力,降低群体茎秆倒伏率和子粒含水量,但遗传方差降低较快。育种可考虑先运用混合选择法改良群体,克服遗传方差迅速降低的缺点,然后进行2～3轮的S1选择法进一步提高产量、适应性及耐自交能力,进而创造优异的育种新材料。     

二穗短柄草2C型蛋白磷酸基因BdPP2C1的克隆及表达分析

从二穗短柄草中扩增得到一个2C型蛋白磷酸酶基因,命名为BdPP2C1。BdPP2C1与拟南芥中PP2C家族进行系统进化分析,结果表明,BdPP2C1与拟南芥A类PP2C家族成员的亲缘关系较近。利用实时荧光定量PCR表达分析系统,分析在非生物逆境胁迫及相关信号分子处理下BdPP2C1基因的表达情况。结果表明:该基因的表达受脱落酸(ABA)和双氧水抑制,并且受渗透和低温胁迫诱导。因此,BdPP2C1是非生物逆境胁迫中的一个应答因子,并且可能受相关信号分子调控。     

YS与1B/1R类型K型小麦雄性不育系配合力的比较研究

为了明确YS与1B/1R两类K型小麦雄性不育系的配合力差异,利用1B/1R类型K型小麦雄性不育系K3314A(A1)和YS型小麦温敏雄性不育系A732(A2)、A3017(A3)作母本与7个恢复系进行不完全双列杂交(NCII),对所得F1代的株高、穗长等7个农艺性状的配合力进行了分析.结果表明,10个亲本各性状的一般配合力效应值(GCA)和各组合特殊配合力效应值(SCA)均具有正向和负向两类, 它们在一定基因型均值的基础上表现为加强或削弱该性状的作用.21个杂交组合中,各性状的特殊配合力最高值都出现在2个YS型温敏雄性不育系组配的组合中.其中A3017/新乡9408、A732/新乡9408和A732/温6有较高的生产利用价值.一般配合力效应值(GCA)在同一性状不同亲本间和同一亲本不同性状间均存在显著差异,2个YS型不育系在产量性状上GCA均表现为正值,K3314A的GCA表现为负值.亲本中A732、K460较为理想.     

香蕉枯萎病菌2个生理小种MAPK基因的克隆及序列分析

为了解香蕉枯萎病菌(Fusarium oxysporum f.sp.cubense)1号生理小种(race 1)和4号生理小种(race4)丝裂原活化蛋白激酶(MAPK)基因focMK1,focMK4的序列差异,根据番茄枯萎病(Fusarium oxysporum f.sp.lycopersici)的MAPK基因相关序列设计引物,利用PCR和RT-PCR技术,分别克隆了香蕉枯萎病菌2个生理小种的基因序列和开放阅读框。结果表明,所获得的2个MAPK基因均含有3个内含子和4个外显子,2个基因序列只有一个碱基差异,编码氨基酸355个,氨基酸组成无差异;根据生物信息学软件预测编码蛋白没有信号肽,具有2个功能位点,分子量和pI分别为41ku和6.47。该基因编码的蛋白具有高度保守性,与大多数植物致病菌的同源性都在90%以上,不同病原镰刀菌间的同源性接近100%。     

SSR Analysis on Diversity of AA Genome Oryza Species in the Southeast and South Asia

To investigate genetic diversities among the AA genome Oryza species in the Southeast and South Asia, a total of 428 accessions of the AA genome Oryza species were genotyped using 36 simple sequence repeats (SSR) markers distributed throughout the rice genome. All of the 36 SSR markers generated polymorphic bands, revealing 100% polymorphism. The number of alleles per locus ranged from 3 to 17 with the mean of 8.6. The Nei’s genetic diversity index (He) ranged from 0.337 at RM455 to 0.865 at RM169 with an a...     

Grain Quality and Genetic Analysis of Hybrids Derived from Different Ecological Types in Japonica Rice （Oryza sativa）

The performance and inheritance of 7 quality traits were studied using F2 rice grain derived from 8×8 diallel crossing made by employing 8 parents of different ecological japonica rice types. Differences in each trait among 8 parents were not obvious, but in F2 generation, transgressive phenomena were found in all the traits studied, indicating that the genes controlling these traits among parents were segregated. The inheritance of grain width, grain weight, chalkiness score (CS), gelatinization temperature (GT) and gel consistency (GC) were suitable to additive-dominant model, and dominant effect contributed mainly for the 5 traits. The inheritance of grain length (GL) and amylose content (AC) did not fit into additive-dominant model, existing epistatic interactions. Dominant genes for grain width and grain weight had the efficiency of decreasing effect, and dominant genes for CS, GT and GC had the efficiency of enhancing effect. Koshihikari contained more recessive genes for gelatinization temperature than other varieties. Zhendao 88 had more dominant genes in grain width and grain weight than other varieties. Xiushui 04 possessed more dominant genes for GL and GC, and more recessive genes for CS than other varieties.     

不同人工老化法测定玉米种子耐储性的比较分析

以耐储玉米自交系东156和不耐储自交系东237为试材,对40℃高温高湿老化法、饱和盐加速老化法、58℃±1℃热水浴老化法和50%甲醇老化法4种玉米种子人工老化方法进行比较,筛选适合玉米种子耐储性评价的方法。结果表明,经过人工老化处理后,东156与东237种子耐储性相关指标均有所下降。4种老化方法中只有热水浴老化法处理后,东156的发芽势、发芽率、发芽指数、活力指数、相对发芽势、相对发芽率、相对发芽指数和寿命指数等耐储性相关指标均显著或极显著高于东237,其中寿命指数差异最大。