广东特殊生境的普通野生稻（Oryza rufipogon Griff）遗传变异研究

选择来自广东佛冈县地处山林区远离稻田的一个普通野生稻生境中的25份野生稻样本,以国外多年生、一年生普通野生稻7份样本和广东地方栽培稻8个品种为对照进行种植观察,调查了21个形态生物学性状,对调查数据作方差分析表明,有20个性状表现极显著差异,1个性状表现显著差异。通过模糊聚类分析,可将25份样本分为4类,各类与对照材料之间表现不同的遗传差异,说明该生境野生稻存在较丰富的遗传变异,但从形态生物学和生育期观察,从中尚未发现一年生型、栽培型、粳型等变异类型的分化。     

广东高州普通野生稻(Oryza rufipogon Griff)的遗传多样性和居群遗传分化研究

利用30对SSR引物对广东高州普通野生稻3个群体进行了遗传多样性分析和居群遗传分化研究.结果表明,30对引物中只有20对表现出多态,多态位点比率P为66.7%;在20个多态位点中共检测出81个等位变异,平均等位变异数(Ap)为4.05 个;3个群体总的遗传多样性(Ht)为0.61,其中,居群内的遗传多样性为居群间的遗传多样性的3倍多,说明总的遗传多样性主要来自居群内;虽然居群间的遗传分化系数(G ST)较低,仅为0.2427,但当遗传相似系数临界值增加时,3个群体在聚类图上相对独立 ,说明3个群体既存在着高度的遗传相似性,又具有一定程度的遗传分化,可以作为3个居群进行原生境保护.     

广西普通野生稻(Oryza rufipogon Griff)表型性状和SSR多样性研究

以中国普通野生稻初级核心种质中广西普通野生稻部分中的 2 2 3份野生稻为材料 ,以平均分布于水稻 12条染色体上的 34对SSR引物和中国稻种资源目录中的表型性状分析广西普通野生稻SSR位点的等位变异、多样性的地理分布及不同生长习性间的多样性分布等。结果表明 ,每对引物检测到的多态性片段 7～ 4 8条 ,平均为 2 4 .91条 ,普通野生稻的等位变异数明显大于地方稻种 ,在所分析的SSR位点中杂合位点比例变化在 1.35 %～ 81.31%之间 ,平均为 32 .0 1% ,与自花授粉的栽培稻相比具有较高的杂合率 ;北纬 2 2°～ 2 3°和 2 3°～ 2 4°范围内的两个区域内(一个包括隆安、扶绥和邕宁三县 ,另一个包括象州、来宾、武宣、玉林和贵港五个县 )所包含的普通野生稻数量多 ,遗传多样性大 ,在DNA水平上是广西普通野生稻的遗传多样性中心 ,而表型性状多样性中心是在北纬 2 1°～ 2 2°和2 2°～ 2 3°,其多样性分布与DNA水平不完全一致。在 4种生长习性间 ,DNA水平上的遗传多样性大小依次为匍匐型 ,倾斜型 ,半直立型和直立型 ,表型水平的多样性与DNA水平的多样性基本一致。     

普通野生稻（Oryza rufipogon Griff．）的SSR遗传多样性研究

利用SSR标记对分布在我国7个省的17个居群普通野生稻(Oryza rufipogon Griff．)的种质资源进行研究,结果表明：17个居群普通野生稻的遗传多样性有着显著的差异,从UPGMA聚类结果可以看出普通野生稻的遗传多样性与其生态地理分布有显著的相关性,遗传多样性指数高的地区极有可能是栽培稻的起源中心;用筛选出的7对SSR引物对336份：DNA样品进行扩增,得到60条特异条带,在分子水平上进一步证明普通野生稻的起源为两广地区;本研究也表明SSR是进行遗传资源多样性研究的一种切实有效的研究方法。     

普通野生稻（Oryza rufipogon）DNA导入栽培稻后代主要性状的遗传变异

通过花粉管通道法将普通野生稻(Oryza rufipogon)DNA导入宁夏水稻品种宁粳16号和宁粳23号中,获得外源DNA导入系(以D表示),选择的导入系经4次连续自交获得D4代材料.选择其中36个株系在北京种植,进行农艺性状调查和变异类型分析.结果表明,这些导入系分别在株型、生育期、分蘖力、株高、每穗粒数、每穗粒重、穗长、千粒重等性状上发生变异;一些导入系的每穗实粒数、每穗粒重、穗长、千粒重和分蘖数等明显高于受体,表现出高产潜力.本文着重阐述导入系的每穗实粒数、千粒重、分蘖数等与产量相关性状的遗传变异.     

云南元江普通野生稻(Oryza urfipogon Griff)遗传多样性分析及保护策略研究

用SSR方法对云南元江普通野生稻3个自然居群进行30个位点的遗传多样性分析.结果表明,元江普通野生稻具有较高的遗传变异水平(Ap＝2.6,Hs=0.77),且群体遗传分化系数较大,GST为41.08%,即在遗传变异总量中41.08%存在于居群间.本文还通过对云南元江普通野生稻遗传多样性特点的分析,提出了保护策略.     

广东高州普通野生稻稻瘟病抗性鉴定

在苗期采用人工接种稻瘟病混合菌株对108份高州普通野生稻样本进行了稻瘟病抗性鉴定,经过初鉴和复鉴,筛选出抗级为2—3级的抗病样本3份;利用我国稻瘟病菌5个菌群7个生理小种22个菌株对初鉴表现中抗的2份种茎样本B35和C1的各2个F1单株进行苗期抗谱测定,获得全群抗性频率分别达到86．36％和81．82％的材料各1份。本鉴定结果可供高州野生稻资源的进一步深入研究和开发利用参考。     

广东高州普通野生稻的研究进展

广东高州普通野生稻具有丰富的遗传多样性,是水稻育种的宝贵资源。本文就近几年来有关高州普通野生稻遗传多样性、生殖特性、有利基因发掘与利用等方面的研究进行综述,并提出进一步研究的设想。     

广东普通野生稻调查、收集与保护建议

为了全面了解广东省普通野生稻自然生存现状,更好地为我国野生稻资源的长远保护提供决策依据,2005-2016年我们对广东普通野生稻自然生存状况进行了实地调查。结果查明,目前广东共有25个县(市)尚存普通野生稻,尚有分布点118个,其中103个有历史资料记载,15个为本次调查新发现。根据历史资料统计,广东省原有普通野生稻分布点1083个,但截至目前野生稻已全部消失的分布点有980个,分布点丧失率90.49%,呈现严重濒危的趋势。调查发现,造成野生稻大量消失的原因主要有垦荒造田、水利建设、城镇建设、养殖业发展、除草剂使用、环境污染和外来物种侵袭等。调查的同时,从118个点共收集了1371份种茎样本进行异位保存,为普通野生稻资源持续利用提供了更大的物质保障。此外,对广东普通野生稻的长远保护提出了建议。     

SSR 在琼海和三亚两普通野生稻居群中的遗传变异

通过分析37个SSR座位在琼海与三亚两普通野生稻(Oryza rufipogon)居群中的遗传变异, 结果表明, SSR座位在三亚普通野生稻居群中的变异高于其在琼海普通野生稻居群中的变异。根据遗传相似性和遗传距离公式得到琼海与三亚普通野生稻居群间的遗传相似性为0.6385, 遗传距离为0.4486 cM。Wright的FST检验结果表明, 这37个SSR座位在两居群之间存在着中等程度的遗传分化, FST=0.3909。此分化结果主要是由两居群间弱的基因漂移导致的(Nm=0.3895)。     

Allozyme variation and population genetic structure of common wild rice Oryza rufipogon Griff. in China

In order to determine the genetic diversity and genetic structure of populations in common wild rice Oryza rufipogon, an endangered species, allozyme diversity was analyzed using 22 loci in 607 individuals of 21 natural populations from the Guangxi, Guangdong, Hainan, Yunnan, Hunan, Jiangxi and Fujian provinces in China. The populations studied showed a moderate allozyme variability (A=1.33, P=22.7%, Ho=0.033 and He=0.068), which was relatively high for the genus Oryza. The levels of genetic diversity for Guangxi and Guangdong were significantly higher than those for the other regions, and thus South China appeared to be the center of genetic diversity of O. rufipogon in China. A moderate genetic differentiation (F_ST=0.310, I=0.964) was found among the populations studied. Interestingly, the pattern of population differentiation does not correspond to geographic distance. An estimate of the outcrossing rate (t=0.324) suggests that the species has a typical mixed-mating system. The deficit of heterozygotes (F=0.511) indicates that some inbreeding may have taken place in outcrossing asexual populations because of intra-clone outcrossing events and ”isolation by distance” as a result of human disturbance. In order to predict the long-term genetic survival of fragmented populations, further studies on gene flow among the remaining populations and the genetic effects of fragmentation are proposed. Finally, some implications for the conservation of endangered species are suggested. Received: 22 June 1999 / Accepted: 20 December 1999     

Plant regeneration from protoplasts of wild rice (Oryza rufipogon Griff.)

Embryogenic callus initiated from basal segments of micropropagated shoots of Oryza rufipogon were used to initiate cell suspension cultures. After approximately 3 months these cultures were capable of yielding large numbers of protoplasts which underwent sustained division in agarose-solidified medium at a frequency comparable to that observed with Japonica rice protoplasts in previous studies. O. rufipogon plants were reproducibly regenerated from the protoplast-derived callus and are currently being grown to maturity. This is the first report of plant regeneration from protoplasts of a wild species of Oryza.Abbreviations BAP 6-benzyalamino purine - 2,4-D 2,4-dichlorophenoxyacetic acid - MES 2[N-morpholino] ethanesulphonic acid - NAA -naphthalene acetic acid - PAR photosynthetically active radiation - SCV settled-cell volume     

Identification of the blast resistance gene Picl(t) from Chaling common wild rice (Oryza rufipogon Griff.)

Rice blast, caused by the fungal pathogen Magnaporthe oryzae (M. oryzae), is one of the most destructive and widespread plant diseases in the world. Utilization of resistance genes in rice breeding is considered to be an effective and economical method to control this disease. To identify new sources of blast resistance, a set of 1160 introgression lines (ILs) containing chromosome segments of Chaling common wild rice (Oryza rufipogon Griff.) in the genetic background of an elite indica rice variety 93-11 were developed and phenotyped in the blast nursery. Thirty-three ILs displaying stable blast resistance in three consecutive years were obtained. Among them, one line, IL1043, was subsequently found to be resistant to all of the 28 M. oryzae isolates from different regions through artificial inoculation in greenhouse. By combining bulk segregant analysis coupled with next-generation sequencing (BSA-seq) and recessive class analysis (RCA), a major blast resistance gene in IL1043, designated Picl(t), was mapped on rice chromosome 6 flanked by the markers RM527 and Indel6 with an interval of approximately 925 kb, which covers the Pi2/9 locus. These results will facilitate fine mapping and cloning of Picl(t), and the linked markers will further provide a useful tool for rice blast resistance breeding.     

Analysis of QTLs for yield-related traits in Yuanjiang common wild rice （Oryza rufipogon Griff.）

Using an accession of common wild rice(Oryza rufipogon Griff.)collected from Yunnjiang County,Yunnan Province,China,as the donor and an elite cnltivar 93-11,widely used in two-line indica hybrid rice production in China,as the recurrent parent,an advanced backcross populations were developed.Through genotyping of 187 SSR markers and investigation of six yield-related traits of two generations(BC4F2 and BC4F4),a total of 26 QTLs were detected by employing single point analysis and interval mapping in both generations.Of the 26 QTLs,the alleles of 10(38.5%)QTLs originating from O.rufipogon had shown a beneficial effect for yield-related traits in the 93-11 genetic background.In addition,five QTLs controlling yield and its components were newly identified,indicating that there arc potentially novel alleles in Yuanjiang common wild rice.Three regions underling significant QTLs for several yield-related traits were detected on chromosome 1,7 and 12.The QTL clusters were founded and corresponding agronomic traits of those QTLs showed highly significant correlation,suggesting the pleiotropism or tight linkage.Fine-mapping and cloning of these yield-related QTLs from wild rice would be helpful to elucidating molecular mechanism of rice domestication and rice breeding in the future.     

Analysis of QTLs for yield-related traits in Yuanjiang common wild rice (Oryza rufipogon Griff.)

Using an accession of common wild rice (Oryza rufipogon Griff.) collected from Yuanjiang County, Yunnan Province, China, as the donor and an elite cultivar 93-11, widely used in two-line indica hybrid rice production in China, as the recurrent parent, an advanced backcross populations were developed. Through genotyping of 187 SSR markers and investigation of six yield-related traits of two gen- erations (BC4F2 and BC4F4), a total of 26 QTLs were detected by employing single point analysis and inte...     

Assessment of population genetic structure in common wild rice Oryza rufipogon Griff. using microsatellite and allozyme markers

The genetic structure of five natural populations of common wild rice Oryza rufipogon Griff. from China, was investigated with 21 microsatellite loci and compared to estimates of genetic diversity and genetic differentiation detected by 22 allozyme loci. Microsatellite loci, as expected, have much higher levels of genetic diversity (mean values of A = 3.1, P = 73.3%, Ho = 0.358 and He = 0.345) than allozyme loci (mean values of A = 1.2, P = 12.7%, Ho = 0.020 and He = 0.030). Genetic differentiation detected by microsatellite loci ( FST = 0.468, mean I = 0.472) was higher than that for allozyme loci ( FST =0.388, mean I = 0.976). However, microsatellite markers showed less deviation from Hardy-Weinberg expectation (Wright's inbreeding coefficient FIS = -0.069) than do allozymes ( FIS = 0.337). These results suggest that microsatellite markers are powerful high-resolution tools for the accurate assessment of important parameters in population biology and conservation genetics of O. rufipogon, and offer advantages over allozyme markers.     

东乡野生稻回交重组系的抗旱性评价体系

选用协青早B//东乡野生稻/协青早B回交重组系及其亲本共79份水稻材料,研究了水稻发芽期、苗期、孕穗期、成熟期的种子萌发抗旱指数等31个指标与东乡野生稻回交重组系抗旱性的关系.结果表明:15％ PEG-6000溶液处理的相对发芽势、20％ PEG-6000溶液处理的种子萌发抗旱指数和相对发芽势及水分胁迫后最长根长、苗高、根鲜质量、根干质量、根系相对含水量、萎蔫率、叶片可溶性糖含量、叶片脯氨酸含量、叶片丙二醛含量、叶片相对含水量、卷叶级别、株高、单株分蘖数、单株有效穗数、穗实粒数、着粒密度、结实率、千粒重等21个指标的相对值与抗旱系数或抗旱指数的相关性显著.通过逐步回归分析筛选出9个抗旱性鉴定指标:20％ PEG-6000溶液处理的相对发芽势及水分胁迫后根干质量、根系相对含水量、叶片可溶性糖含量、叶片脯氨酸含量、叶片丙二醛含量、单株有效穗、结实率和千粒重的相对值.根据这些指标和偏相关系数,建立了不同时期的抗旱性评价(D值)方程和评价体系,可对东乡野生稻回交重组系抗旱性进行较好的评价.     

Comparison of the genetic diversity of common wild rice (Oryza rufipogon Griff.) and cultivated rice (O. sativa L.) using RFLP markers

Forty fourth single-copy RFLP markers were used to evaluate the genetic diversity of 122 accessions of common wild rice (CWR, Oryza rufipogon Griff.) and 75 entries of cultivated rice (Oryza sativa L. ) from more than ten Asian countries. A comparison of the parameters showing genetic diversity, including the percentage of polymorphic loci (P), the average number of alleles per locus (A), the number of genotypes (Ng), the average heterozygosity (Ho) and the average genetic multiplicity (Hs) of CWR and indica and japonica subspecies of cultivated rice from different countries and regions, indicated that CWR from China possesses the highest genetic diversity, followed by CWR from South Asia and Southeast Asia. The genetic diversity of CWR from India is the second highest. Although the average gene diversity (Hs)of the South Asian CWR is higher than that of the Southeast Asian CWR, its percentage of polymorphic loci (P), number of alleles (Na) and number of genotypes (Ng) are all smaller. It was also found that the genetic diversity of cultivated rice is obviously lower than that of CWR. At the 44 loci investigated, the number of polymorphic loci of cultivated rice is only 3/4 that of CWR, while the number of alleles, 60%, and the number of genotypes is about 1/2 that of CWR. Of the two subspecies studied, the genetic diversity of indica is higher than that of japonica. The average heterozygosity of the Chinese CWR is the highest among all the entries studied. The average heterozygosity of CWR is about two-times that of cultivated rice. It is suggested that during the course of evolution from wild rice to cultivated rice, many alleles were lost through natural and human selection, leading to the lower heterozygosity and genetic diversity of the cultivated rice. Received: 19 May 1999 / Accepted: 26 April 2000     

Fine mapping of a quantitative trait locus for grain number per panicle from wild rice (Oryza rufipogon Griff.)

SIL040, an introgression line (IL) developed by introgressing chromosomal segments from an accession of Oryza rufipogon into an indica cultivar Guichao 2, showed significantly less grains per panicle than the recurrent parent Guichao 2. Quantitative trait locus (QTL) analysis in F₂ and F₃ generations derived from the cross between SIL040 and Guichao 2 revealed that gpa7, a QTL located on the short arm of chromosome 7, was responsible of this variation. Alleles from O. rufipogon decreased grains per panicle. To fine mapping of gpa7, a high-resolution map with 1,966 F₂ plants derived from the cross between SIL040 and Guichao 2 using markers flanking gpa7 was constructed, and detailed quantitative evaluation of the structure of main panicle of each of F₃ families derived from recombinants screened was performed. By two-step substitution mapping, gpa7 was finally narrowed down to a 35-kb region that contains five predicted genes in cultivated rice. The fact that QTLs for five panicle traits (length of panicle, primary branches per panicle, secondary branches per panicle, grains on primary branches and grains on secondary branches) were all mapped in the same interval as that for gpa7 suggested that this locus was associated with panicle structure, showing pleiotropic effects. The characterizing of panicle structure of IL SIL040 further revealed that, during the domestication from common wild allele to cultivated rice one at gpa7, not only the number of branches and grains per panicle increased significantly, more importantly, but also the ratio of secondary branches per panicle to total branches per panicle and the ratio of grains on secondary branches per panicle to total grains per panicle increased significantly. All these results reinforced the idea that gpa7 might play an important role in the regulation of grain number per panicle and the ratio of secondary branches per panicle during the domestication of rice panicle.Feng Tian and Zuo Feng Zhu contributed equally to this work.