野生稻与栽培稻及种间杂种F1叶表面亚显微结构的研究

用扫描电镜对原产中国的３种野生稻、２个栽培稻品种及栽培稻与野生稻间的杂种Ｆ１,就气孔频度、气孔器乳突、大瘤状乳突及本栓细胞乳突等叶片表面亚显微结构作了比较观察研究。结果显示这些性状具有种的特性,发现种间杂种的亚星微性状与两个亲本 特性及亲缘关系远近有关。     

稻属叶表皮结构特征及其系统学意义

本文对稻属Oryza L. 23种植物叶片表皮的结构特征进行了观察和研究,结果表明:叶表皮的某 些性状,如在叶片脉带之间长细胞中乳突的大小和分布以及叶表皮气孔器上小乳突的数目和着生位置 在稻属的各种之间有着一定的变异规律,这在稻属各种的分类和其系统关系的研究中有一定的价值。 综合这些性状的变异特征,按照叶片下表皮脉间长细胞中乳突的大小和分布特征以及气孔器中小乳突 的数目和着生位置可将稻属这23个物种分为3组。第一组包括长颖野稻、马来野稻、疣粒野稻和颗粒 野稻,这些种的叶表皮脉间长细胞中没有大乳突和中乳突,仅偶见极稀疏分布的小乳突,气孔器中均无 小乳突。第二组包括短药野稻、二倍体和四倍体药用野稻、小粒野稻、紧穗野稻、斑点野稻、阔叶野稻、高 株野稻、大颖野稻、根茎野稻和澳洲野稻,这些种的叶表皮脉间长细胞中通常没有大、中乳突,但密布小 乳突,且大多数种的气孔器保卫细胞的近两端各有2个小乳突。第三组包括栽培稻、一年生普通野稻、 多年生普通野稻、长雄蕊野稻、展颖野稻、南方野稻、矮舌野稻、非洲栽培稻和希来特野稻,这些种的叶 表皮脉间的长细胞中常有大乳突、中乳突和小乳突,而气孔器保卫细胞的近两端各有2个明显的小乳 突,并同时常在气孔器副卫细胞的近内缘还有2～4个小乳突。     

中国三种野生稻谷粒外稃乳突结构的电镜观察

用扫描电镜对中国的普通野生稻、药用野生稻、疣粒野生稻以及栽培籼、粳稻谷粒外稃的乳突结构进行了观察和比较。结果表明：普通野生稻、药用野生稻和栽培籼稻的乳突属锐型双峰乳突,疣粒野生稻的乳突属瘤型单峰乳突,栽培粳稻属钝型双峰乳突。谷粒外稃乳突特征是稳定的微观形态形状。     

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     

云南野生稻叶茎根组织结构特性的比较研究

采用徒手切片法对云南3种野生稻和栽培稻的叶片、茎秆及根的组织结构进行比较研究,以明确野生稻的内部结构,为进一步揭示其结构与云南野生稻的生长势旺盛、营养吸收能力强、抗某些病虫害能力强的关系奠定基础。结果表明,(1)云南野生稻与栽培稻叶片主脉、茎秆及根的组织结构差异显著,其中景洪疣粒野生稻、景洪药用野生稻与栽培稻的差异最明显。(2)在叶主脉结构中,景洪疣粒野生稻无气腔结构,维管束数量少、面积小;景洪药用野生稻、3个普通野生稻材料存在多个维管束和气腔结构,维管束、束内导管直径及气腔面积较栽培稻大,而栽培稻中的气腔均为2个。(3)在茎秆结构方面,景洪疣粒野生稻茎秆最小,维管束数量最少,其茎壁内的维管束排列方式与栽培稻不同;景洪药用野生稻和普通野生稻茎秆及茎壁较栽培稻粗厚,维管束数量也较栽培稻多,普通野生稻的茎壁中有通气组织。(4)在根的组织结构中,3种野生稻的导管数量较多,导管直径及中柱面积较栽培稻大,外皮层出现了具有凯氏带功能的凯氏点等。     

栽培稻与疣粒野生稻杂种F1代的基因组原位杂交鉴定

生物素标记的疣粒野生稻总DNA作探针,未标记的栽培稻总DNA封阻,对栽培稻与疣粒野生稻杂种F1体细胞染色体进行基因组原位杂交（Genomic in situ hybridization,简称GISH）分析。FITC检测表明,杂种细胞中来自瘛发粒野生稻的染色体有较多的黄色或黄绿色荧光信号,来自栽培稻的染色体只检出很少的信号。每条疣粒野生稻染色体上信号点所占的总的区域只是染色体的一小部分,表明疣粒野生稻染色体与栽培稻染色体的DNA序列大部分是同源的。     

药用野生稻和栽培稻挥发油的化学成分比较研究

本文采用水蒸气蒸馏法提取抗褐飞虱品种药用野生稻(Oryza officinalis)和感褐飞虱品种栽培稻(O.sativa)的挥发油,并运用气相色谱-质谱法(GC/MS)对二者的化学成分进行种类和含量的比较分析。结果表明,两种植物材料的挥发油在种类和含量上存在明显差异,如,糠醛(药用野生稻7.60%栽培稻0%;2,3-苯并二氢呋喃(药用野生稻10.80%,栽培稻3.32%);4-乙烯基-2-甲氧苯酚(药用野生稻22.99%,栽培稻7.32%)。推测含量较高的糠醛、2,3-苯并二氢呋喃和4-乙烯基-2-甲氧苯酚可能是药用野生稻对褐飞虱的活性成份。     

野生稻和栽培稻的光合作用和叶绿素含量的比较研究

对三个野生稻种,即普通野生稻(Oryza sativa f. spontanea)、药用野生稻(O.officinalis)、疣粒野生稻(O. meyeriana),一个由野生稻演变为栽培稻的过渡型深水莲,五个栽培稻(O.sativa L.)地方品种(六月谷、饭萝白、矮仔占、夜花占、桂161),和三个栽培稻改良品种(包选二、珍珠矮、广陆矮)的光合作用和叶绿素含量作比较研究。结果表明,野生稻的光合作用光饱和点比栽培稻的低;前者为25000～35000lx,后者为35000～45000lx。在饱和光强下,野生稻的光合作用也比栽培稻弱,前者平均为13.0mg CO_2 dm~(-2)h~(-1),后者平均为20.6mg CO_2 dm~(-2)h~(-1)。深水莲的光合作用强度和光饱和点均大约介于野生稻和栽培稻之间。 野生稻的叶绿素含量和叶绿素a/b比亦比栽培稻低;前者平均分别为1.93mg g~(-1) FW和2.55,后者平均分别为2.83 mg g~(-1)FW和3.12。深水莲亦有较高的叶绿素含量和叶绿素a/b比。 上述这些性状,是和野生稻原来是分布在比较荫蔽的生态环境,因而具有阴生植物的光合性状,而栽培稻则比较地具有阳生植物的性状相一致的。     

稻属三种植物叶片下表皮的形态比较

稻属有20多种,其中3种和其他物种明显不同.Oryza eichingeri被认为在亚洲和非洲间断分布;O. officinalis和O punctata被证明同时含有二倍体和四倍体.为了识别种内微形态差异,利用光镜和扫描电镜,对这3个物种的成熟叶片下表皮进行了观察.结果表明,O. eichingeri的叶脉长细胞乳突形状在种内是不一致的,除了斯里兰卡地理宗具有的瘤状乳突外,乌千达地理宗还具有长条状的乳突;同样,种内多样性也存在于O. officinalis和O.punctata内,特别存在于它们不同倍性的细胞型间.在D officinalis中,四倍体有粒状气孔器外围乳突和脉带表皮毛较少,二倍体细胞型具有三角形拱状排列的气孔器外围乳突和较密集的脉带表皮毛;而在O. punctata中,和四倍体相比,二倍体具有小的长细胞表面乳突和低密度分布的气孔.这些稳定的差异为3种植物的种内鉴定和分类修订提供了新的证据.     

稻属单拷贝核基因TPI序列分析及其在疣粒野生稻鉴定中的应用

通过对稻属(Oryza L.)及其近缘属等18个禾本科植物单拷贝核基因TPI序列进行扩增和测序,分析其系统进化关系及序列差异,并设计鉴定疣粒野生稻(O.granulata)特异的分子标记.序列分析表明,栽培稻间TPI基因序列碱基变异少,野生稻相对变异丰富,其中疣粒野生稻变异尤为明显,根据疣粒野生稻的TPI基因序列特异位点设计引物,利用这一分子标记对疣粒野生稻进行准确鉴定.     

The Structural Features of Leaf Epidermis in Oryza and Their Systematic Significance

The rice genus (Oryza L. ) belongs to the grass family(Poaceae) and contains 24 annual or perennial species, including two cultivated rice species, i.e., the Asian rice ( O. sativa L. ) and African rice (O. glaberrima Steud. ), and 22 wild species distributed throughout the tropics of the world. Species in this genus have been extensively studied by scientists with different approaches, including morphological characterization and cytological and molecular investigations. The leaf epidermis is an important morphological character which has been studied for taxonomic identification and studies on systematic relationships of species, particularly in grasses. In this study, morphological features of the leaf epidermis of 23 rice species were observed through light microscopy. The results showed that some characters of the rice leaf epidermis had significant diversity between species and these characters were valuable for the identifying Oryza species, and for assessing systematic relationships in the genus. For example, O.schlechteri, O.ridleyi, O.longiglumis, O.granulata, and O. rneyeriana had elliptic stomatal complexes, whereas the other species had rhombic stomatal complexes. In most cases, papillae on the surface of the epidermis were variable in size and distribution between species. The size of papillae varied from small ( 1.5～4.4µm in diameter), medium-sized (9～18µm), to large (21～30µm) , and the pattern of papillary size and distribution were very useful for identification of rice species. In addition, the number and location of the small papillae in stomatal complexes were particularly different between species. Based on the following combinations of leaf-epidermic characters, i.e., the size and distribution of papillae on the abaxial surface of the epidermis, the number and location of the small papillae in stomatal complexes, and the shape of stomatal complexes, the 23 studied Oryza species could be divided into three major groups. The first group comprises O.longiglumis, O.ridleyi, O.meyeriana, and O.granulata. In these species, neither large nor medium-sized papillae, in some cases extremely rare small papillae, were found on the surfaces of epidermis, and there were no small papillae found in stomatal complexes. All species in the first group had elliptic stomatal complexes. The second group consists of O.brachyantha, diploid and tetraploid O.officinalis, O.minuta, O.eichingeri, O. punctata, O.latifolia, O.alta, O.grandiglumis, O.rhizomatis, and O.australiensis. In these species usually no large papillae were observed, but medium-sized and densely populated small papillae were found to cover the surface of epidermis, and at least four small papillae were found in stomatal complexes (in guard cells) of most species. The third group contains O.sativa, O.nivara, O.rufipogon, O.longistaminata, O. glumaepatula, O.meridionalis, O.barthii, O.glaberrima and O. schlechteri. The abaxial leaf epidermis of these species was usually covered with large papillae, medium-sized, and small papillae. In addition, more than 4 (usually 6～8 ) small papillae were found in guard cells or/and subsidiary cells of the stomatal complexes. Most species in the second and third groups had rhombic stomatal complexes. These results agree mostly with previous re-ports on the biosystematic studies of rice species by applying other methodologies.     

云南野生稻不同染色体组型和外植体材料的离体培养研究

云南野生稻不同外植体愈伤组织诱导能力差别较大。花粉培养中愈伤组织诱导率差异在0％～11．8％之间,用成熟胚诱导愈伤组织,其诱导率在18．0％～35．2％之间,茎叶培养则在12．0％～25．0％之间。云南野生稻不同外植体诱导的愈伤组织再分化为绿苗的分化率在8．3％～100．0％之间。疣粒稻组培特性最好,东乡普通野生稻和景洪普通野生稻次之,药用稻最难组培。本文建立了疣粒、东乡、景洪普野3种野生稻的离体无性系,为长期保存云南野生稻资源奠定了基础。     

Characterization of Interspecific Hybrids Between Oryza sativa L. and Three Wild Rice Species of China by Genomic In Situ Hybridization

In the genus Oryza, interspecific hybrids are useful bridges for transferring the desired genes from wild species to cultivated rice （Oryza sativa L.）. In the present study, hybrids between O. sativa （AA genome） and three Chinese wild rices, namely O. rufipogon （AA genome）, O. officinalis （CC genome）, and O. meyeriana （GG genome）, were produced. Agricultural traits of the F1 hybrids surveyed were intermediate between their parents and appreciably resembled wild rice parents. Except for the O. sativa × O. rufipogon hybrid, the other F1 hybrids were completely sterile. Genomic in situ hybridization （GISH） was used for hybrid verification. Wild rice genomic DNAs were used as probes and cultivated rice DNA was used as a block. With the exception of O. rufipogon chromosomes, this method distinguished the other two wild rice and cultivated rice chromosomes at the stage of mitotic metaphase with different blocking ratios. The results suggest that a more distant phylogenetic relationship exists between O. meyeriana and O. sativa and that O. rufipogon and O. sativa share a high degree of sequence homology. The average mitotic chromosome length of O. officinalis and O. meyeriana was 1.25- and 1.51-fold that of O. sativa, respectively. 4＇,6＇-Diamidino- 2-phenylindole staining showed that the chromosomes of O. officinalis and O. meyeriana harbored more heterochromatin, suggesting that the C and G genomes were amplified with repetitive sequences compared with the A genome. Although chromocenters formed by chromatin compaction were detected with wild rice-specific signals corresponding to the C and G genomes in discrete domains of the F1 hybrid interphase nuclei, the size and number of O. meyeriana chromocenters were bigger and greater than those of O. officinalis. The present results provide an important understanding of the genomic relationships and a tool for the transfer of useful genes from three native wild rice species in China to cultivars.     

Inoculation with Xanthomonas oryzae pv. oryzae induces thylakoid membrane association of Rubisco activase in Oryza meyeriana

Oryza meyeriana is a wild species of rice with high resistance to Xanthomonas oryzae pv. oryzae (Xoo), but the resistance mechanism is poorly understood. Protein gel blot analysis and immuno-gold electron microscopy showed that Xoo infection induced an association of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) activase (RCA) with the thylakoid membrane in O. meyeriana, which led to considerable decline in the initial activity and the activation state of Rubisco. In susceptible cultivated rice, RCA remained in the chloroplast stroma. RCA may play a role in resistance to Xoo in O. meyeriana that differs from its well-known role in activating Rubisco, perhaps by protecting the thylakoid membrane against damage from Xoo.     

Somatic cell cryopreservation and protoplast regeneration of important disease-resistant wild rice Oryza meyeriana Baill

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稻属植物胚的形态结构与二（异）型子叶

长久以来植物学界认定稻 (OryzasativaL .)是单子叶植物。作者从稻胚发育的研究中确认稻胚具二型子叶 ,并非单子叶。稻属其他种的胚胎形态与O .sativa是否相同 ?是否具二型子叶 ?根据扫描电子显微镜的观察结果 ,稻属 (Oryza) 2 2个种和亚种的胚的形态结构可以分为两种类型。O .sativa等 16个种胚具腹鳞和侧鳞 ,属第一类型 ;O .meyeriana (Zoll.etMor.exSteud .)Baill.ssp .tuberculataW .C .WuetY .G .Lu ,G .C .Wang等 6个种 (亚种 )胚缺腹鳞和侧鳞 ,属第二类型。O .sativa和O .meyerianassp .tuberculata的胚胎发育过程所出现的盾片原基、胚根鞘原基、胚芽鞘原基和生长锥均来自原胚 ,前二者发育成胚套 ,是外围子叶 ;胚芽鞘原基发育成围在生长锥外并盖住生长锥的空心的倒锥状胚芽鞘 ,是顶生子叶。第一类型与第二类型稻胚都具有二型子叶。第二类型稻胚在盾片原基发育过程中并不分化出腹鳞和侧鳞 ,因而造成第二类型稻胚缺腹鳞与侧鳞。稻的二型子叶源于原胚的背腹极性分化     

云南野生稻中的抗白叶枯病基因Xa21的鉴定

水稻白叶枯病是水稻生产上的主要细菌病害之一。从野生稻中发掘优异的水稻白叶枯病抗性材料,可以拓宽栽培稻抗白叶枯病遗传基础。经过温室接菌鉴定和PCR标记分析,对云南野生稻进行Xa21基因的检测鉴定。温室接菌鉴定表明,云南野生稻对广谱致病小种PX099及云南强致病菌Y8具有较好的抗性能力,特别是疣粒野生稻对致病菌株达到免疫程度;PCR标记分析表明,云南野生稻不含有Xa21基因,但含有与Xa21基因某些区域同源的片段。本研究结果为寻找新的抗源材料及快速发掘利用云南野生稻中的抗白叶枯病基因提供理论依据。     

野生稻基因组随机扩增多态性DNA(RAPD)分析

用18个随机引物对2份栽培稻、12份包含有六个基因组型的野生稻DNA进行了扩增,共获得147个多态性DNA片断,把这些多态性DNA片断作为遗传位点用UPGMA法计算出各材料间的遗传相似性系数,并作了聚类分析．主要结果如下：1普通野生稻同栽培稻的亲缘关系很近,其中江永普通野生稻更接近于粳稻．2．CCDD组的Oryzalatifolia和EE组的O．australiensis遗传多态性相似。3．B、C、D、E组的遗传多态性相似,组成一个复合体,此复合体与A组的遗传多态性也相似,而F组则相距较远．4．O．mcyeriana和Rhynchofyzasabulata尚未确定组型,RAPD测定结果表明,前者与其它组型的种亲缘关系较远,后者则与AC复合体的种较近．     

A biosystematic study of theOryza meyeriana complex (Poaceae)

To determine the taxonomic status and to clarify the biosystematic relationships of wild rice species, i.e.,Oryza granulata andO. meyeriana in theO. meyeriana complex (Poaceae), a series of intraspecific and interspecific hybridization experiments, genomic analyses through meiotic pairing, and morphological studies were conducted. Hybridization results demonstrate comparatively high crossability with about 34–39% of seed set, and very limited reproductive isolation betweenO. granulata andO. meyeriana. Chromosome pairing in the parental species and their F₁ hybrids indicate very high genomic affinity between the two taxa. Morphological analyses of the parental accessions and their intra- and interspecific hybrids show a large variation among the samples included, but no appreciable grouping was observed. The traditionally used character, i.e., spikelet length, does not contribute significantly to the grouping of the samples included. Based on the facts that the two taxa have negligible reproductive barriers and share almost identical genomes, the authors believe thatO. granulata andO. meyeriana do not warrant separate taxonomic status, and recommend that they be combined asOryza granulata.