利用SRAP标记研究海南野生稻的遗传多样性与遗传分化

利用8对多态性较好的SRAP引物对海南120份普通野生稻、55份疣粒野生稻和26份药用野生稻进行扩增,在检测到的219个位点中,普通野生稻的多态性位点率为74.89％,疣粒野生稻为42.47％,药用野生稻25.11%。香农指数以普通野生稻最高0.3277,疣粒野生稻为0.2044,药用野生稻最低0.1113。UPGMA聚类分析结果显示供试材料与地理来源相一致,相关性强,各居群个体间没有出现任何交叉。根据居群间的遗传分化系数,普通野生稻群体的基因多样性为0.2135,群体内的平均基因多样性大于居群间的基因漂变,说明普通野生稻居群遗传分化不显著,遗传多样性主要来自于居群内,基于群体杂合度和居群遗传多样性指数特点,认为实施保护策略时,优先保护遗传多样性最丰富的WDL和WDA居群。疣粒野生稻居群存在中等程度的遗传分化,建议原生境保护;药用野生稻居群数量较少,建议原生境保护。     

海南野生稻资源调查收集与保护

为了加强海南野生稻资源的保护和研究利用,2002—2013年对海南省18市(县)野生稻资源进行野外调查、资源收集和原、异位保护。初步查明,目前在全省15个市(县)发现154个野生稻自然居群,其中普通野生稻136个、疣粒野生稻11个、药用野生稻7个。收集到野生稻居群87个,其中普通野生稻80个、疣粒野生稻4个、药用野生稻3个,总共2900余份野生稻种茎,经繁殖和编目后妥善保存于海南省农业科学院试验基地的热带野生稻异位保存圃中,并参照农业行业标准-农业野生植物原生境保护点建设技术规范,分别在文昌、琼海、儋州、万宁、陵水和保亭等县(市)建立了6个野生稻原生境保护点,其中普通野生稻4个,疣粒野生稻1个,药用野生稻1个。根据调查结果,对海南野生稻濒危状况及其根源进行了分析,提出了海南野生稻保护相关建议。     

中国野生稻的现状调查

中国有三种野生稻。野生稻是栽培水稻的野生近缘种,已被列为濒危植物^[7],它们分布于中国南方８个省（区）的热带、亚热带湿热生境中。 近年来人为的干扰和生境的破坏已导致它们的居群大量绝灭,其中濒危程度最高的是普通野生稻。本文阐述了它们的生物学、生态学特性,研究了它们的群落学特征,初步报道了它们的濒危状况,探讨了致濒原因。     

居群遗传结构研究中显性标记数据方法初探

为对比显性标记应用于居群遗传结构研究时不同统计参数的适用性,利用RAPD技术对中国5个居群的100个疣粒野生稻个体进行了遗传结构分析。在衡量居群遗传多样性水平时,多态位点比率（PPB）会低估遗传变异的量,其价值不如Shannon多样性指数和Nei基因多样性指数,而采用Nei指数时不必进行Lynch-Milligan矫正。对个体间遗传关系进行分析时,17种遗传相似性指数矩阵两两之间的Mantel检测都表现出极显著的相关性（r＞0.95,t＞t     

云南疣粒野生稻的居群遗传结构及其在原位保护中的意义

用 1 2种酶系统 1 7个等位酶位点对采自云南思茅地区疣粒野生稻Oryzameyeriana(Zoll.etMerr.exSteud .)Baill.7个居群的 1 6 4个个体进行了遗传多样性的研究 .相对于以前报道过的具有相同生活史特性和繁育系统的种子植物和稻属中的其他种类 ,疣粒野生稻的遗传多样性水平极低 (A =1 .1 ,P =8.0 % ,H_o=0 .0 0 4和H_e=0 .0 1 5 ) ,但居群间的遗传分化较大 .F_ST高达 0 .6 4 9,表明在遗传变异总量中的6 4 .9%存在于居群间 .对这样在较小的地理区域内居群间就呈现出剧烈遗传分化的物种来说 ,大多居群均有保护的价值 .因此 ,进行原位保护时原则上应建立较大的自然保护区 ,涉及较多数目的居群 .同时 ,提出了在云南进行原位保护时应优先考虑的地点 .     

用组织培养法繁殖疣粒野生稻研究

疣粒野生稻是优异的种质资源,由于其种子的强休眠性,常规方法很难繁殖成功,本实验利用组织培养方法,通过诱导试管苗和诱导愈伤再分化成苗两种途径,对疣粒野生稻的试管微繁进行了研究,获得了一定数量的疣粒野生稻试管苗及成熟种子。本研究结果表明,疣粒野生稻直接成苗及愈伤分化成苗对培养基的选择性均不强,用组织培养法快速繁殖疣粒野生稻是行之有效的途径。     

中国野生稻资源及其抗虫性

野生稻在中国有3种：普通野生稻（Oryza rufipogon Griff.),药用野生稻（O.officinalis Watt.et Watt)和疣粒野生稻（O.meyeriana Baill)。主要分布在热带亚热带的江河流域。多项研究表明,野生稻对多种害虫有较强的抗性,利用野生稻的优良特性采用杂交技术选育出新的抗虫品种,或从野生稻中提取抗虫物质对野生稻的利用和环境保护有重要意义。     

云南疣粒野生稻稻瘟病抗性

野生稻(Oryza rufipogo)保存有许多栽培稻(O. sativa)不具备或已经消失的优异基因资源, 是扩大栽培稻遗传背景、改良产量与品质、提高抗病虫害及抗逆境能力的重要基因库。疣粒野生稻(O. meyeriana)是中国3种野生稻资源之一, 主要分布在云南。为进一步了解其稻瘟病抗性, 首先利用来自不同稻作区的稻瘟病菌株, 通过注射接种法对疣粒野生稻进行系统的稻瘟病抗性鉴定, 发现疣粒野生稻对接种的所有稻瘟病菌株都感病。进一步采用3'/5' RACE方法, 从疣粒野生稻中克隆了水稻同源基因Pid2和Pid3, 并构建过表达转基因株系对基因功能进行了研究。结果表明, Pid2和Pid3与疣粒野生稻中同源基因间在DNA和氨基酸水平上有较大的序列差异, 过表达转基因的日本晴植株对稻瘟病菌的敏感性与对照相似。推测疣粒野生稻在自然接种条件下, 表现出的抗稻瘟病表型很可能是其旱生叶片结构特征形成了对稻瘟病菌侵染的天然屏障。对控制疣粒野生稻这一类性状基因资源的挖掘和利用, 有利于优良抗性水稻品种的培育。研究结果为疣粒野生稻的研究利用提供了新信息和新思路。     

中国野生稻的种类、地理分布及其特征特性综述

野生稻在世界上主要分布于热带和亚热带。由于所处生态环境的复杂性 ,野生稻在其漫长的进化过程中 ,形成了极其丰富的遗传多样性 ,是宝贵的种质资源 ,是发展稻作育种的重要物质基础。中国南方地处热带、亚热带 ,气候炎热 ,夏日长暖 ,雨量充沛 ,十分适宜野生稻的生长与繁衍 ,野生稻分布很广 ,资源十分丰富。据调查 ,我国有 3种野生稻 ,即普通野生稻、药用野生稻和瘤粒野稻 (中国“疣粒野生稻”)。我国野生稻分布广泛 ,南自海南省三亚 ,北至江西省东乡 ;东自台湾省桃园 ,西至云南省盈江。纬度约跨 1 0度 ,经度约跨 2 3度。中国野生稻的特征特…     

我国北回归线区域普通野生稻遗传多样性和遗传结构研究

选择分布于水稻染色体组的26对SSR引物,对集中分布于广西来宾市北回归线附近的13个普通野生稻居群的342份材料进行遗传多样性和遗传结构研究。结果表明,该地区的野生稻无论在种内(A=9.154,Ae=4.446,I=1.547,He=0.671)还是居群水平上(P=95%,A=4.219,Ae=2.394,I=0.905,He=0.476)遗传多样性都十分丰富,高于同类研究水平。供试居群的遗传分化系数Gst为0.30,表明30%的遗传变异存在于居群间,大部分遗传变异存在于居群内。居群间的遗传一致度变化范围为0.332~0.903,且居群间地理距离越近,遗传一致度越高,说明北回归线附近的普通野生稻居群符合"隔离-距离"模型。综合上述研究结果和我国野生稻保护现状,建议对居群G13和G06实施优先保护。     

Distribution and Conservation of an Endangered Wild Rice Oryza granulata in China

Nearly Twenty years after the first large-scale field survey on wild rice in China during 1978-1982, we have recently made an extensive survey on Chinese Oryza granulata (Nees et Arn. Ex Watt.), including its current distribution, population characteristics and status of endangerment. Because of the change in the division of administrative areas, the number of counties or cities where the wild rice grows has reached up to 30. Oryza granulata grows normally under the coverage of 90%-210% in the tropical forest with resistance to drought and moderate disturbance. The measurement of aggregation showed that it maintains a colony pattern in the community. Oryza granulata scatters seeds mainly by gravity and animals, causing limited scale of gene flow between local populations and attaining a typical metapopulation structure of the distributional pattern. Our survey revealed that 12.9% of the populations had been extinct, and 83.9% were endangered. The extent of disturbance varied in different areas. Up to date, the majority of the populations grow in the mountainous areas of southwestern Hainan and drainage area of Nanding river, Lancang river of Yunnan, under the threat of human activity. A total DNA bank was established that consists of 1109 individuals from 49 populations. In the practice of conservation, difficulties such as lacking immediate utility and scattered distribution impeded the conservation efficient. However, owing to its characteristics of population genetics, sustainable and ex situ conservation strategies are appropriate in some places to salvage this important rice germplasm.     

六个核基因片段在中国颗粒野生稻中的单核苷酸多态性及其在群体遗传学研究中的应用

为了筛选可用于亚洲栽培稻的重要野生近缘物种颗粒野生稻群体遗传结构研究的有效SNPs标记,本研究共选取Osgstf3、OsNAC6、BADH、XCP2、EMF2和ERCC4 6个重要水稻功能基因,检测了来自中国云南和海南12份颗粒野生稻代表样品中的单核苷酸多态性,结果发现Osgstf3、OsNAC6、BADH和XCP2的单核苷酸多态性接近于零,但在EMF2和ERCC4内含子区中检测到较低的核苷酸变异。利用EMF2和ERCC4片段对来自云南的3个天然群体和来自海南的2个天然群体共52个个体进行了群体遗传学研究,发现该物种具有高水平的遗传分化和低水平的遗传变异的独特群体遗传结构;这种剧烈的遗传分化不仅来自云南和海南两个地区之间,而且还来自地区内居群间,说明该野生稻生长的热带、亚热带森林生境的破坏导致了碎裂的居群间基因流强烈受阻。     

Analysis of genetic variability of South American wild rice populations (Oryza glumaepatula) with isozymes and RAPD markers

The knowledge of population structure and genetic diversity of wild relatives of rice is needed to investigate their evolutionary history and potential use in breeding programs. Very little is known about the wild rice species ( Oryza spp.), particularly those that are native to South America. A study using isozyme and RAPD markers was conducted to estimate the level of genetic diversity of four South American wild rice populations ( Oryza glumaepatula ) recently collected in the Amazon forest and western Brazil rivers. F -statistics and genetic diversity parameters calculated from isozyme and RAPD markers indicated high values for inbreeding coefficients and differentiation among the four populations. In agreement with this, a pattern of greater variation between than within populations was observed with both types of markers. These findings were corroborated by an AMOVA analysis, which indicated that a large portion of the total genetic variation was attributed to regional divergence. The partition of the AMOVA analysis among populations showed that most of the genetic diversity was due to differences among populations. This distribution pattern of genetic variation of O. glumaepatula populations is in agreement with the expectation for an autogamous species and provides important baseline data for conservation and collection strategies for this species.     

原、异位保存东乡野生稻主要农艺性状的比较研究

为了解原、异位保存东乡野生稻主要农艺性状的变化,对庵家山普通野生稻原、异位群体进行了考察和比较分析。结果表明,原、异位保存的东乡野生稻在每穗总粒数、每穗实粒数、结实率等方面存在较大的差异,如异位保存的东乡野生稻每穗总粒数平均为58.28粒,比原位多11.44粒,但其每穗实粒数却较少,为9.89粒,结实率降低了45.43%。从种植环境可以发现,异位保存圃的环境不适宜东乡野生稻的生长,应该选择与原生境相类似的种植环境,以便于东乡野生稻的保存、研究与利用。     

海南省万宁市野生荔枝资源种群特征研究

野生荔枝是荔枝(Litchi chinensis Sonn.)进行品种改良与创新的重要基因库,该研究通过对海南万宁自然保护区内、外野生荔枝资源进行实地调查,并用空间分布格局、种群径级结构、静态生命表、存活曲线等方法进行比较分析,以揭示野生荔枝种群在保护区内、外的分布规律及其种群动态趋势,为野生荔枝资源的保护提供依据。结果表明:(1)海南万宁自然保护区内、外野生荔枝种群均为聚集分布,且非自然保护区聚集程度比自然保护区的聚集程度更为明显。(2)径级结构分析发现,自然保护区内野生荔枝种群结构为增长型,种群生命期望值整体上基本随径级增加呈递减趋势,存活曲线呈近直线型,说明自然保护区内的野生荔枝种群处于相对稳定的状态;而自然保护区外的野生荔枝种群为衰退型。(3)群动态预测结果也显示,自然保护区内外的野生荔枝种群的动态变化指数(Vpi′)以及随机干扰风险极大值均趋于零,但自然保护区外的值均大于区内,说明保护区外受到的敏感性比自然保护区内更强,即保护区外野生荔枝种群的生存环境已受到严重的人为干扰威胁。研究认为,在自然保护区外,野生荔枝种群幼树的严重缺失不仅与野生荔枝种群自身的更新机制有关,更重要的是生存环境的破坏和人为砍伐导致野生荔枝种群结构发生变化,野生荔枝林面积下降。     

Population Ecological Characteristics of the Rare and Endangered Plant Camellia rhytidophylla from Guizhou

Camellia rhytidophylla is endemic and endangered and with important economic species, from the ⅢD10d in eastern Asia, which distributed at 573-920m in evergreen broad leaved deciduous mixed forest in Luowang river valley of Kaiyang county. So, it is significant to study the population characteristics, conservation and utilization. We took general survey in suspected distribution area about 6 square kilometers, set up plots in dense area, and analyzed the population structure, development and spatial distribution pattern. The results showed that the structure of Crhytidophylla populations were increasing and the proportion of young tree in population was 4638%. The size of population density was young shrubs＞middle aged shrubs＞adult shrubs. The survival curve of population was Deevey Ⅲ model. There were 2 peaks in the Ⅰand Ⅳ age classes on the mortality rate curve and disappearance rate curve respectively, then there are 2 troughs in the Ⅲ and Ⅵ age classed at the same time. The spatial distribution pattern of Crhytidophylla significantly differentiated at different stages of development, the young individuals were aggregated at all spatial scales while the middle aged individuals were aggregated at small spatial scales and randomly distributed at larger scales. Differences in the distribution of adult individuals could be attributed to artificial disturbance and habitat heterogeneity. The spatial pattern was not close in different stages of development, which all showed negative or no correlation. The spatial distribution pattern of Crhytidophylla was the interaction of the factors, such as its biological characteristics,habitat heterogeneity, and artificial disturbance, etc.. The key factor limiting population development was low natural reproduction rate; the primary factors causing its endemic distribution mainly included habitat heterogeneity, topographical constraints on seed dispersal and artificial disturbance.     

Diversity in the Content of Some Nutritional Components in Husked Seeds of Three Wild Rice Species and Rice Varieties in Yunnan Province of China

In addition to rice (Oryza sativa L.) cultivars, there are three wild rice species, namely O.rufipogon Griff, O. officinalis Wall and O. granulata Baill, in Yunnan Province, China. Each species has different subtypes and ecological distributions. Yunnan wild rice species are excellent genetic resources for developing new rice cultivars. The nutritional components of the husked seeds of wild rice have not been investigated thus far. Herein, we report on the contents of total protein, starch, amylose, 17 amino acids, and five macro and five trace mineral elements in husked seeds from three wild rice species and six O. sativa cultivars. The mean (± SD) protein content in the husked rice of O. rufipogon, O. officinalis, and O. granulata was (14.5 ± 0.6)%, (16.3 ± 1. 1)%, and (15.3 ± 0.5)%, respectively. O. officinalis Ⅲ originating from Gengma had the highest protein content (19.3%). In contrast, the average protein content of six O. sativa cultivars was only 9.15%. The total content of 17 amino acids of three wild rice species was 30%-50% higher than that of the six cultivars. Tyrosine, lysine, and valine content in the three wild rice species was 34%-209% higher than that of the cultivars. However, the difference in total starch content among different O. sativa varieties or types of wild rice species was very small. The average amylose content of O. rufipogon, O. officinalis,and O. granulata was 12.0%, 9.7%, and 11.3%, respectively, much lower than that of the indica and japonica varieties (14.37%-17.17%) but much higher than that of the glutinous rice cultivars (3.89%). The sulfur, phosphorus, magnesium, zinc, and ferrite content in the three wild rice species was 30%-158% higher than that of the six cultivars. The considerable difference in some nutritional components among wild rice species and O. sativa cultivars represents a wide biodiversity of Yunnan Oryza species. Based on the results of the present study, it is predicted that some good genetic traits, especially high protein and ideal amylose content, of Yunnan wild rice species may be useful in improving the nutritional value of rice. This is the first report regarding the amino acid, mineral element, protein and amylose content of husked seeds of some Yunnan wild rice species that have important genetic characteristics for rice quality and nutritional value.     

ITS1 Sequences of Nuclear Ribosomal DNA in Wild Rices and Cultivated Rices of China and Their Phylogenetic Implications

he first internal transcribed spacer (ITS1) of nuclear ribosomal DNA of three wild rice species and two subspecies of cultivated rice, which are distributed in China, was amplified using PCR technique and sequenced with automated fluorescent sequencing. The sequences of ITS1 ranged from 193 bp to 218 bp in size and G/C content varied from 69.3%to 72.7%. In pairwise comparison among the five taxa, sequence site divergence ranged from 1.5 % to 10.6%. Phylogenetic analysis of ITS1 sequences using Wagner parsimony generated a single well-resolved tree, which revealed that Oryza rufipogon was much more closely related to cultivated rice species than to the other two wild species. Oryza granulata was less closely related to either cultivated rice species or the other two wild species, and might be a unique and isolated taxon in the genus Oryza. The phylogenetic relationships of the three wild rice species and two cultivated rice subspecies inferred from ITS1 sequences is highly concordant with those based on the molecular evidence from isozyme, chloroplast DNA (cpDNA), mitochondrial DNA (mtDNA) and nuclear DNA (nDNA) of the genus Oryza.     

中国野生稻及栽培稻核糖体DNA第一转录间隔区序列分析及其系统学意义

用 PCR技术从产于我国的 3种野生稻和亚洲栽培稻的 2个亚种中特异地扩增和测序了 r DNA的第一转录间隔区。普通野生稻 (Oryza rufipogon)、药用野生稻 (O.officinalis)、疣粒野生稻 (O.granu-lata)和栽培稻的两个亚种 (O.sativa ssp.indica,O.sativa ssp.japonica)的 ITS1序列为 1 93bp、1 94bp、2 1 8bp、1 94bp和 1 94bp,它们的 G/ C含量为 69.3%～ 72 .7% ,序列中位点趋异率为 1 .5%～ 1 0 .6%。序列的相似性比较和简约性分支分析的结果表明 ,普通野生稻与栽培稻的两个亚种之间的亲缘关系最为密切 ;药用野生稻与普通野生稻和与栽培稻的两个亚种的相似性都为 82 % ,说明它与 AA基因组有一定的亲缘关系 ;疣粒野生稻与普通野生稻、药用野生稻和栽培稻两个亚种的亲缘关系相对较远 ,它在稻属中可能是一个系统地位较独特的类群。以 ITS1序列构建的 3种野生稻和 2个栽培稻亚种的系统发育关系与前人用同工酶、叶绿体 DNA、线粒体 DNA和核 DNA资料重建的稻属的系统发育关系基本一致