Physical localization of the 18S-5.8S-26S rDNA and sequence analysis of ITS regions in Thinopyrum ponticum (Poaceae: Triticeae): implications for concerted evolution

Fluorescence in situ hybridization was used in Thinopyrum ponticum, a decaploid species, and its related diploid species, to investigate the distribution of the 18S-5.8S-26S rDNA. The distribution of rDNA was similar in all three diploid species (Th. bessarabicum, Th. elongatum and Pseudoroegneria stipifolia). Two pairs of loci were observed in each somatic cell at metaphase and interphase. One pair was located near the terminal end and the other in the interstitial regions of the short arms of one pair of chromosomes. However, all of the major loci in Th. ponticum were located on the terminal end of the short arms of chromosomes, and one chromosome had only one major locus. The maximum number of major loci detected on metaphase spreads was 20, which was the sum of that of its progenitors. The interstitial loci that exist in the possible diploid genome donor species were probably 'lost' during the evolutionary process of the decaploid species. A number of minor loci were also detected on whole regions of two pairs of homologous chromosomes. These results suggested that the position of rDNA loci in the Triticeae might be changeable rather than fixed. Positional changes of 18S-5.8S-26S rDNA loci between Th. ponticum and its candidate genome donors indicate that it is almost impossible to find a genome in the polyploid species that is completely identical to that of its diploid donors. The possible evolutionary significance of the distribution of the rDNA is also discussed. Internal transcribed spacer (ITS) regions of nuclear DNA in Th. ponticum were investigated by PCR amplification and sequencing. The sequence data from five positive clones selected at random, together with restriction site analysis, indicated that the ITS repeated units are nearly homogeneous in this autoallodecapolypoid species. Combined with in situ hybridization results, the data led to the conclusion that the ITS region has experienced interlocus as well as intralocus concerted evolution. Phylogenetic analyses showed that the sequences from Th. ponticum have concerted to the E genome repeat type.     

普通小麦-百萨偃麦草(Thinopyrum bessarabicum)二体异附加系的选育与鉴定

为转移与利用百萨偃麦草耐盐、抗病等优良基因,用普通小麦中国春-百萨偃麦草双倍体与中国春杂交,通过染色体C-分带、分子原位杂交并结合减数分裂中期I的染色体配对分析,从回交后代中选育出一套小麦-百萨偃麦草二体异附加系。对这套异附加系进行的鉴定与分析表明,各附加系除添加了一对百萨偃麦草染色体外,小麦的21对染色体未见明显变化。各附加系所添加的百萨偃麦草染色体在减数分裂中期I配对基本正常,仅有少量单价体,其自交后代中外源染色体亦能正常传递。这说明所培育的这套二体异附加系在细胞学上已相对稳定,暂分别编号为DAJ1、DAJ2、DAJ3、DAJ4、DAJ5、DAJ6和DAJ7。各异附加系中百萨偃麦草染色体在小麦族中的部分同源群归属和百萨偃麦草耐盐抗病基因在染色体上的定位研究正在进行之中。     

大赖草及近缘物种原位杂交和Southern杂交的分析

用Ｔｈｉｎｏｐｙｒｕｍｂｅｓａｒａｂｉｃｕｍ（Ｓａｖｕｌ．＆Ｒａｙｓ）Ａ．Ｌｖｅ的基因组ＤＮＡ作探针,分别与大赖草Ｌｅｙｍｕｓｒａｃｅｍｏｓｕｓ（Ｌａｍ．）Ｔｚｖｅｌ．和脆轴偃麦草Ｔｈ．ｊｕｎｃｅｕｍ（Ｓａｖｕｌ．＆Ｒａｙｓ）Ａ．Ｌｖｅ的体细胞杂交,大赖草的１４对染色体均出现杂交信号,脆轴偃麦草只有７对染色体有杂交信号。在用重复ＤＮＡ序列ＰＨｖ６２作探针的原位杂交中,Ｔｈ．ｂｅｓａｒａｂｉｃｕｍ有４对染色体有杂交信号,大赖草有１３对染色体显示杂交信号,新麦草Ｐｓａｔｈｙｒｏｓｔａｃｈｙｓｊｕｎｃｅａ（Ｆｉｓｃｈ．）Ｎｅｖｓｋｉ和脆轴偃麦草无杂交信号。用ＰＨｖ６２作探针的Ｓｏｕｔｈｅｒｎ杂交结果与原位杂交相似。在被检测的１２个普通小麦大赖草异源染色体系中,除二体附加系中５Ｌｒ＃１和双二体附加系１Ｌｒ＃１＋５Ｌｒ＃１没有杂交信号外,其余的异染色体系与ＰＨｖ６２都有特异杂交信号。据此推测Ｔｈ．ｂｅｓａｒａｂｉｃｕｍ有可能参予了赖草属物种的形成过程。但是,大赖草的染色体组在进化过程中显然已发生过变异。     

大赖草及近缘种染色体C—分带的研究

对Ｔｈｉｎｏｐｙｒｕｍｎｂｅｓａｒａｂｉｃｕｍ（Ｓａｖｕｌ．＆Ｒａｙｓ）Ａ．Ｌｖｅ、新麦草（Ｐｓａｔｈｙｒｏｓｔａｃｈｙｓｊｕｎｃｅａ（Ｆｉｓｃｈ．）Ｎｅｖｓｋｉ）和大赖草（Ｌｅｙｍｕｓｒａｃｅｍｏｓｕｓ（Ｌａｍ．）Ｔｚｖｅｌ．）染色体Ｃ分带的核型进行了比较研究。Ｔｈ．ｂｅｓａｒａｂｉｃｕｍ和新麦草的染色体在Ｃ分带带型上有明显的差异,显示了物种的特异性。３个物种的核型表明,Ｃ带带纹主要分布在染色体的末端,大部分染色体不显着丝粒带和中间带。在大赖草染色体上的末端带很明显。一些大赖草的染色体具有与Ｔｈ．ｂｅｓａｒａｂｉｃｕｍ和新麦草某些染色体相似的Ｃ带带型。对大赖草染色体组与Ｔｈ．ｂｅｓａｒａｂｉｃｕｍ和新麦草染色体组的关系进行了讨论。     

THINOPYRUM BESSARABICUM和THINO-PYRUM ELONGATUM的基因组关系研究

对 2个八倍体 C.S- Thinopyrum bessarabicum( AABBDDJJ,2 n=8x=56)和 Goshawk( GHK) - Thinopyrum elongatum( AABBDDEE,2 n=8x=56)的根尖细胞染色体进行 C-分带 ,从中分检出 Th.bessarabicum和 Th.elongatum的各自染色体进行核型分析 ,结果表明 :Th.bessarabicum和 Th.elongatum的大多数染色体都具有端带 ,但 Th.bessarabicum的端带更强 ,很少有中间带 ;而 Th.elongatum的染色体除 E1外其它染色体的端带较弱 ,而且带纹较丰富 ,有较多的中间带。对 C.S- Th.bessarabicum和 GHK- Th.elongatum进行有性杂交 ,其杂交种F1的 PMC染色体在 MI的平均配对构型为 1 5.50 5.0 3 [ 1 4.2 0 ○ 0 .40 0 .2 1 ,其中 Th.bessarabicum和 Th.elongatum染色体平均配对成 2 .5个二价体 ,由 C-分带显示大多数 Th.bessarabicum和 Th.elongatum染色体呈单价体状态。因此推断 ,Th.bessarabicum和 Th.elongatum两物种的染色体应属不同的染色体组     

Short-term effects of aluminium at alkaline pH on the structure and function of the photosynthetic apparatus

A 24 h exposure of the salt-tolerant grass Thinopyrum bessarabicum (Savul. and Rayss) A. Love seedlings to 1 mM aluminium (Al) in nutrient solution at pH of 9.0 resulted in a significant reduction of the biomass. In control samples the mesophyll chloroplasts exhibited the usual lens shape with most grana arranged in straight or slightly curving lines, and only 6.5 % of the grana were out of order. In Al-treated plants the mesophyll chloroplasts displayed a slightly distorted shape and distended size with most grana arranged in bow-like lines, while in the central region of the organelle as many as 26.7 % of the grana were independent and out of order in relation to the long axis. The morphological changes in the chloroplast shape and grana arrangement were probably due to swelling and distension of the chloroplasts in consequence to the altered membrane permeability. The initial in vivo chlorophyll (Chl) fluorescence FO, as well as the intermediate FI and peak fluorescence FP were increased under the Al stress: this indicated a destruction of photosystem (PS) 2 reaction centres and increased reduction of QA. The (FI-FO)/(FP-FO) ratio exhibited a significant increase indicating higher proportion of PS2 centres unable to reduce QB. Changes in the chloroplast ultrastructure seemed to be the reason of photosynthetic electron transport inhibition. Yet all these changes in the photosynthetic performance and chloroplast ultrastructure were considered as indirect effects of Al treatment since Al concentration in the leaves was undetectable. Disturbances in the chloroplast ultrastructure could be caused by a reduced uptake and/or transport of other nutrients. This revised version was published online in June 2006 with corrections to the Cover Date.     

Fluorescence in situ Hybridization and Southern Blot Analyses on Leymus racemosus and Related Species

Fluorescence in situ hybridization (FISH) was carried out in somatic cells of racemosus (lam.) Tzvel. and Thinopyrum junceum ( Savul. & Rayss) A. Love using Th. bessarabicum ( Savul. & Rayss) A. Love genomic DNA as probe. Fourteen pairs of chromosomes in L. racemosus gave positive signal, and only seven of fourteen pairs of chromosomes in Th. junceum showed signal. In FISH probed with PHv62, the chromosomes in Th. bessarabicum and L. racemosus hybridized with PHv62, whereas Th. Bessarabicum showed positive signal in four pairs of chromosomes, and the latter in thirteen pairs of chromosomes. No positive signal was observed in chromosomes of Psathyrostachys juncea (Fisch.) Nevski and Th. junceum. The results of Southern hybridization probed with PHv62 were similar to those of in situ hybridization. Twelve alien chromosome lines of T. aestivum-L, racemosus were detected by PHv62 probing. In most of the alien chromosome lines, PHv62 hybridized in fragment of Leymus chromosomes, except for the line containing chromosomes 5Lr # 1 and 1 1Lr # 1. It is inferred that Th. bessarabicum may involve in the formation of species. Non the less, significant changes have occurred during the evolution of Leymus genomes.