Real-time PCR is a closed DNA amplification system that skillfully integrates biochemical, photoelectric and computer techniques. Fluorescence data acquired once per cycle provides rapid absolute quantification of initial template copy numbers as PCR products are generated. This technique significantly simplifies and accelerates the process of producing reproducible quantification of nucleic acid molecules. It not only is a sensitive, accurate and rapid quantitative method, but it also provides an easier way to calculate the absolute starting copy number of nucleic acid molecules to be tested. Together with molecular bio-techniques like microarray, real-time PCR will play a very important role in many aspects of molecular life science such as functional gene analysis and disease molecular diagnostics. This review introduces the detailed principles and application of the real-time PCR technique, describes a recently developed system for exact quantification of AUX/IAA genes in Arabidopsis, and discusses the problems with the real-time PCR process.
Tel: 010-62772243; E-mail: Liujy @ mail.tsinghua.edu.cn。
Seeds have been categorized as orthodox, recalcitrant and intermediate seeds according to their dehydration behaviors. Identification of desiccation-tolerance and -sensitivity of seeds is the basis making storage strategy of seeds and long-term conservation of species gene resources. In addition to the inherent characteristics of the species, developmental status of the seeds, dehydration rate, and the conditions under which they are dried and subsequently re-imbibed are very important factors influencing desiccation tolerance of seeds. Survival, electrolyte leakage rate, and germination/growth rate produced by survived seeds are a excellent synthetic parameter when discussing desiccation tolerance of seeds. Desiccation tolerance of seeds is a quantitative feature. The term "critical water content" is incorrect and has caused some confusion in assessment of seed recalcitrance. A new working approach to quantify the degree of seed recalcitrance has been proposed in this paper.
宋松泉 Patricia BERJAK nORMAN pammenter Tobias M. NTULI 傅家瑞
2. Plant Cell Biology Research Unit, School of Life and Enviironmental Sciences, Unixersity of Natal,Durban, 4041 South Africa; 3. 中山大学生命科学学院，广州510275
通讯作者。Tel: 0691-8715474; E-mail: <sqsong@ xtbg.org.cn>。
The species composition and characteristics of mangrove flora in the world were reviewed and discussed. The results suggested that the world''s mangrove plants have 84 species (including 12 varieties) in 24 genera and 16 families. Of which, true mangrove plants have 70 species (including 12 varieties) in 16 genera and 11 families, and semi_mangrove plants 14 species in eight genera and five families. The Eastern Group has 74 species (including 12 varieties) in 18 genera and 14 families, characterized by the genera Aegiceras, Osbornia, Aegialitis, Bruguiera, Ceriops, Kandelia, Scyphiphora and Nypa etc. The Western Group has only 10 species in six genera and five families, characterized by the endemic one-species family, Pelliceraceae, and the genus Laguncularia. The mangrove flora of China is composed of 26 species (including one variety) in 15 genera and 12 families, four of which are endemic. Hainan is most rich in mangrove species, making up about 96.2% of the Chinese total; Guangdong ranks second, making up about 42.3%. It has been demonstrated that Rhizophora stylosa was mistaken for R. mucronata in Taiwan by previous authors.
王伯荪 梁士楚 张炜银 昝启杰
（1. 中山大学生命科学学院，广州510275; 2. 中国科学院植物研究所植被数量生态学重点实验室，北京100091；3. 中国林业科学研究院森林生态与环境保护研究所，北京100091；
摘要：探讨了世界红树林植物区系的种类组成及其特点.确认全世界共有红树植物16科24属84种(含12变种), 其中真红树为11科16属70种(含12变种), 半红树为5科8属14种.东方类群有14科18属74种(含12变种), 以 Aegiceras, Osbornia, Aegialitis, Bruguiera, Ceriops, Kandelia, Scyphiphora 和 Nypa等属为特征;西方类群有5科6属10种, 具有单种特有科皮利西科Pelliceraceae以及Laguncularia属.中国红树植物区系有12科15属26种(含1变种), 其中特有种4种.海南的红树植物种类最为丰富, 占全国的96.2%; 广东次之, 占42.3%.台湾记载的红茄(Rhizophora mucronata)其实为红树榄R. stylose之误.
朱生伟 秦红敏 孙敬三 田颖川
（1. 中国科学院植物研究所光合作用与环境分子生理学院重点实验室，北京100093；2. 北京大学生命科学学院，北京100871；3. 中国科学院微生物研究所，北京100080）
摘要： 用合成的cry1Ac基因与绿色荧光蛋白基因 (GFP)构成融合蛋白基因 ,然后和改造的GNA基因构建双价抗虫基因植物表达载体pBGbfg ,经根癌农杆菌介导转化了烟草。在紫外灯照射下 ,观察到转基因植株叶片中有较强的绿色荧光 ;经抗虫试验、PCR、Southernblot和Westernblot等检测 ,表明该重组植物表达载体能够在转基因植物中有效表达外源基因 ,转基因植株绿色荧光的表型与其抗虫性密切相关。从而成功地建立了以绿色荧光蛋白基因与抗虫基因组成的融合基因转化系统 ,简化了抗虫转基因植物筛选程序 ,有助于快速获得双价抗虫转基因植株。
关键词： cry1Ac-GFP 融合蛋白基因；双抗虫基因；筛选
通讯作者。Tel: 010-62591431-6236; E-mail: sunjs @ ns. ibcas.ac.cn。
通讯作者。Tel: 010-62642577;E-mail: tianyc @ sun.im.ac.cn。
The molecular composition and evolution of the chalcone synthase (CHS) gene family from five species in Camellia(Theaceae) are explored in this study. Sixteen CHS exon 2 from four Camellia species were amplified from total DNA by PCR method. Three sequences of the fifth species in Camellia and two sequences of Glycine max as the designated outgroups were obtained from GenBank. Our results indicated that CHS gene family in Camellia was differentiated to three subfamilies (A, B, C) during the evolutionary history with six groups (A1, A2, A3, B1, B2, C). Among them, only group A2 was possessed by all five species in this study. However, the other five groups were detected only in some species of the plants studied. All members of CHS gene family in this study had high sequence similarity, more than 90% among the members in the same subfamily and more than 78% among different subfamilies at nucleotide level. According to the estimated components of amino acids, the function of CHS genes in Camellia had been diverged. The nucleotide substitutions of the different groups were not identical. Based on phylogenetic analyse inferred from sequences of CHS genes and their deduced amino acid sequences, we concluded that the CHS genes with new function in this genus were evolved either by mutations on several important sites or by accumulation of the mutations after the gene duplication. A further analysis showed that the diversification of CHS genes in Camellia still occurred recently, and the evolutionary models were different to some extant among different species. So we assumed that the different evolutionary models resulted from the impacts of variable environmental elements after the events of speciation.
杨俊波 田 欣 李德铢 郭振华
摘要： 用PCR方法从4种山茶属(Camellia)(山茶科)(Theaceae)植物的总DNA中分别扩增到CHS基因外显子2的部分序列,经克隆、测序得到16个该基因的序列,这些序列与来自GenBank的该属另一种植物的3个序列及作为外类群的大豆(Glycine max (L.) Merr.)的2个序列一起进行分析.研究表明,山茶属CHS基因家族在进化过程中已分化为A、B、C三个亚家族,包括A1、A2、A3、B1、B2、C 等6类不同的基因成员;其中只有A2类成员为全部被研究的5种植物所共有,而其他5类成员只在部分被研究的植物中发现;所有这些CHS成员具有很高的同源性:在核苷酸水平上同一亚家族内基本上高于90%,不同亚家族间也在78%以上.从推测的氨基酸组成看,山茶属内CHS基因的功能已发生了分化,各类成员的碱基替代率有较大差异; 从分子系统发育树和可能的氨基酸组成分析,山茶属具有新功能的基因成员是在经过基因重复后,或是由少数几个位点的突变而成,或是由逐渐积累的突变而形成的.进一步分析认为,该属CHS基因的分化直到近期还在活跃地进行,并且不同种的进化式样有一定的差别,这种不同的进化式样可能是物种形成后受不同环境因素影响而形成的.
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Anomozamites haifanggouensis (Kimura et al.) Zheng et Zhang comb. nov. is a characteristic Bennettitalean plant in Middle Jurassic flora. Only one specimen was collected from Haifanggou Formation (Middle Jurassic) in Daohugou village, Shantou town, Ningcheng County, Nei Mongol Autonomous Region, China. The stem of plant is slender and repeatedly branches as a dichasial system with a fertile shoot in the fork, forming equal and widely divergent branches, on the branches born with the fronds of Anomozamites type. Its reproductive organs consists of some microsporophylls and a few of the bracteoid small leaves (or receptacular leaves). Some scattered bracteoid leaves and microsporophylls were formerly considered by Pan as dicots. Since then, they were proposed to be a non_committal genus Pankuangia and described them as P. haifanggouensisby Kimura et al. In this study, we discover that these bracteoid small leaves and microsporophylls should be of Anomozamites. This discovery provides strong evidence for the “Pankuangia" determination of the taxonomical position.
共生有生殖器官的 Anomozamites 的新发现
郑少林 张立军 巩恩普
（1. 国土资源部沈阳地质矿产研究所，沈阳110032；2. 吉林大学古生物学和地层学研究中心，长春130026；3. 东北大学资源与土木工程学院，沈阳110004; 4. 辽宁省国土资源厅化石保护管理处，沈阳110032）
摘要： 海房沟异羽叶的新联合(Anomozamites haifanggouensis (Kimura et al.) comb. nov.)是辽西和内蒙古东部中侏罗统海房沟组一种特有的本内苏铁类植物.仅发现一块标本(正、反面印痕).标本上保存3枚营养羽叶,它们同几个苞片状的小叶和一些小孢子叶联合在一起,但球果的雌蕊群部分未被保存.标本被收集于内蒙古东部的宁城县山头乡道虎村附近,产于中侏罗统海房沟组.以往在辽西葫芦岛市的南票和白马石乡上三角城等地的同一层位中也曾发现过很多分散保存的苞片状小叶和相似的小孢子叶.它们最初被潘广(1983)认为是一种双子叶的半被子植物(Cycadicotis),并将小孢子叶视为"具皱纹的雌性种囊".后来,又经Kimura等(1994)详细研究,因未找到任何与被子植物有关的证据,将它们归入一个分类位置不明的形态属Pankuangia,并被描述在P. haifanggouensis种名之下.研究结果表明,本文中被研究的标本是同本内苏铁类的Anomozamites异羽叶相连,从而为这些分散保存的生殖器官的确切分类位置的确定提供了有力的证据.
Numerous leaf compressions of Glyptostrobus europaeus (Brongn.) Ung. (Taxodiaceae) are found in Aquitanian (Lower Miocene) lignified clay localities Bolattam and Akzhar in the Southern Turgay, on the right bank of the Dulygaly_Zhilanshik river (Central Kazakhstan). The finely preserved lignified compression remains of leafy shoots make micro_morphological investigation feasible. Comparative studies of the epidermal features both living G. pensilis C. Koch and a new finding of G. europaeus from Early Miocene of Kazakhstan and illustrate their certain difference. The cuticular organization and epidermal features of fossil leaves, which have been compared with these of “the nearest living relative”, G. pensilis, were studied by scanning electron microscopy (SEM) and light microscopy (LM). The fossil leaves of G. europaeus from Kazakhstan are distinguished by stronger, than in living species, G. pensilis, developed “micro-papillae” (“Kristallucken”), visible in SEM as ring-like structures left on the outer surface of cuticle in both nonstomatal and stomatal zones.
It was substantiated by morphological observation, chromosome and DNA identification that the bargene, conferring the resistance to herbicide basta in transgenic Chinese cabbage (Brassica campestris ssp. chinensis var. utilisTsen et Lee), could intrude into the genomes of turnip (B. campestris ssp.rapifera), heading Chinese cabbage (B. campestris ssp. pekinensis) and other non_heading Chinese cabbage (B. campestris ssp. chinensis) in a relatively high frequency while into genome of B. napusat a low frequency via natural cross_pollination in open field. Herbicide resistant hybrids of B. nigra, B. carinata, andB. juncea could be obtained at certain frequency in the greenhouse by hand pollination. At no time did the resistant hybrid plants of transgenic Chinese cabbage with B. oleracea, R. sativus and seven wild weeds, widespread in China, gained in our experiments.
刘凡 赵泓 姚磊 张军杰 曹鸣庆
（1. 北京蔬菜研究中心，北京100089; 北京农业生物技术中心，北京100089）
摘要： 从形态、染色体及分子水平上证实，转基因不结球白菜(Brassica campestris ssp．chinensis var．utilis Tsen et Lee)中编码除草剂Basta抗性的bar基因能在田间条件下，经自然传粉，以较高频率侵入芜菁(B．campestris ssp．rapifera)、结球白菜(B．campestris ssp．peknensis)和不结球白菜(B．campestrs ssp．chnensis)的基因组中，也能少量侵入同属异种的甘蓝型油菜(B.napus)基因组中；在温室人工辅助授粉条件下，除在上述种中的基因漂移率提高外，bar基因尚能以一定频率侵入同属的黑芥(B．nigra)、埃塞俄比亚芥(B．carnata)、芥菜(B．juncea)基因组中，但始终未能得到转基因白菜与结球甘蓝(B．oleracea)、萝卜(R．sativus)的杂种。转基因白菜与十字花科的7种常见杂草经温室人工辅助授粉，也均未得到抗性杂种。
Leaves from three-year-old solar greenhouse nectarine trees (Prunus persica L. var. nectarina Ait. “Zao Hong Yan”) were used as materials in this study. It was the first time that the ultrastructural characteristics of phloem tissues of source leaves were observed and compared in normal and weak light intensities using the transmission electron microscopy. Results showed that the average diameters of companion cells (CC) and sieve elements (SE) of all kinds of veins were bigger in normal than that in weak light intensity, indicating that light could influence the cell development and growth. Dense cytoplasm with abundant mitochondria, endoplasmic reticulums, multivesicular bodies, vesicles and plastids were observed in normal light intensity. On the contrary, CC with small vacuolar structures and few mitochondrias, endoplasmic reticulums were shown in weak light. Misalignment of grana thylakoid margins of nectarine leaves also was seen in weak light. The sieve pores of SEs were obstructed in weak light. Chloroplasts with numerous starch grains and few mitochondrias were noticed in the mesophyll cell (MES) surrounding the bundle sheath in weak light. The storage of starch grains appeared to result from an unbalance between photosynthate production and export of photosynthates. This observation provided a strong support to the point that most leaves export the most of assimilates in the light time. Plasmodesmal densities between SE/CC, CC/PP (phloem parenchyma cell), PP/PP and PP/BSC (bundle-sheath cell) decreased in weak light. Plasmodesmata were observed between CC/SE (NS) (nacreous-walled sieve element), PP/BSC in branch veins in normal light intensity, but not in weak light. Thus apoplasmic pathway may be the main mode of transport of assimilates in weak light, however symplasmic pathway may be the main mode of transport of assimilates in normal light intensity. These results demonstrated that the solar greenhouse nectarine trees could be adapted to the weak light via the ultrastructure variation of phloem tissues of the source leaves.
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This paper describes changes in the structure of buds and vascular cambium as well as in protein content of one-year-old dormant branches of Eucommia ulmoidesOliv. in water-culture conditions. Results confirm the existence of 2 quiescences (Q1, Q2) and 1 rest (R) phases in this tree during the dormancy period. In the R time, the E. ulmoides cambium was unable to reactivate even though the tree was subjected to exogenous IAA, suitable temperature or required luminosity. Furthermore, pistillate trees entered the dormant phase earlier than staminate ones. The protein content in the bark during Q1 increased significantly (P<0.01), but drastically decreased in the R period, before rising up again at the onset of Q2 (P<0.01). Even though the change pattern of protein content was similar, its occurrence is much earlier in staminate than in pistillate plants. SDS_PAGE revealed a “special" protein band of 11.8 kD in the transitional Q1-R-Q2 stages. This “special" protein bands would be related to the cambial dormancy and to the regulation of the Q1-R-Q2 ransition.
Kalima N'KOMA MWANGE 王学文 崔克明
摘要：对杜仲(Eucommia ulmoides Oliv.)一年生枝条休眠中的芽和维管形成层的结构和蛋白质含量进行了研究,探讨了杜仲不同休眠期转化的生理生化机理.结果证实了杜仲树的休眠期包括2个被动休眠期和一个生理休眠期.在生理休眠期,无论用外源IAA处理,还是给予适当的温度和光照,形成层都不能恢复活动.而且,雌株进入各个休眠阶段的时间都比雄株早.树皮中的蛋白质含量在第一被动休眠期间(Q1)显著地逐步增高(P<0.01),进入生理休眠期(R)后,急剧降低,但第二被动休眠期(Q2)开始后又显著升高(P<0.01).杜仲雌雄株树皮中的蛋白质含量变化趋势基本一致,但雄株发生变化的时间比雌株早.SDS-PAGE电泳结果表明,在Q1-R-Q2的转变期出现一11.8 kD的特异蛋白条带.此特异蛋白条带可能与形成层休眠期中各阶段的转变调节有关.
通讯作者。E-mail: ckm @ pku.edu.cn.
Fluctuations of levels of several endogenous plant hormones in isolated rice (Oryza sativa ssp. japonica) embryos during early and mid-embryogenesis and early stages of germination were studied by enzyme-linked immunosorbent assay (ELISA). Embryos were collected at different days after pollination (DAP) and different days after imbibition (DAI) of mature seeds. The contents of gibberellin 1 (GA1), abscisic acid (ABA), isopentenyladenine and isopentenyladenosine (iPAs), zeatin and zeatin riboside (ZRs) were immunochemically assayed. The GA1 level was the highest among all hormones tested. The variations of GA1 levels were opposite with the ABA levels, with some exceptions. During early and mid_embryogenesis, the levels of GA1 and ABA were the highest at 4 DAP. From 8 to 18 DAP, GA1 level declined, whereas the ABA level increased. During germination, GA1 level increased at 2 DAI whereas simultaneously the ABA content decreased. The highest ratio of GA1/ABA was observed at 2 DAI. The levels of iPAs and ZRs were maxima in the embryos at 4 DAP, decreased to a very low level and maintained constant thereafter. Our results provide further evidence that GA1 plays an important role in the early stages of embryo development and germination. The balance between GA1 and ABA, rather than their absolute contents, controls these processes throughout the development, whereas iPAs and ZRs may play important roles in early embryogenesis. The use of isolated embryos as starting material avoids the usual interferences with other tissues such as the endosperm. In addition, this is the first report dealing with the hormonal balance of early-embryos in rice.
杨 军 赵 洁 周 燮 杨弘远
（1. 武汉大学植物发育生物学教育部重点实验室，武汉430072; 南京农业大学农学院，南京210095）
摘要： 以酶联免疫吸附检测技术分析了水稻(Oryza sativa ssp. japonica)分离胚不同发育时期及萌发早期的内源激素含量的动态变化.GA1含量是所测激素中含量最高的.GA1的变化趋势基本上与ABA相反.花后4 d的胚中GA1和ABA的含量最高;花后8 d到18 d,GA1的含量下降,而ABA含量增加.在早期萌发过程中,种子吸涨后2 d的胚中GA1含量迅速上升,而ABA下降.GA1/ABA的最高比值也出现在吸涨后2 d的胚中.iPAs和ZRs的最高含量也出现在开花后4 d的胚中,但随后含量均下降到相当低的水平,并几乎没有变化.研究结果进一步证实了GA1在早期胚胎发生和萌发过程中起重要的作用;推测iPAs和ZRs可能仅在胚胎发生的早期起作用;GA1与ABA含量之间的相对平衡控制着胚胎发育的过程.用分离胚作为测试材料可以避免胚乳等其他组织成分的干扰,从而比较准确地反映了胚的内源激素变化.此外,本研究是首次用4 d的水稻幼胚作为激素含量测定的起始材料.
通讯作者。E-mail: <hyyang 2@ whu.edu.cn>。
A series of new cognitions on the morphogenesis of maize (Zea mays L.) embryo have been obtained with scanning electron microscopy and semi_thin section techniques. 1. The proembryo. The proembryo from zygotic cell divisions may be divided into three parts: proper, hypoblast and suspensor. The suspensor is short and small, and only exists transiently. As to the hypoblast there is a growth belt, which promotes elongation of the hypoblast. Eventually the upper portion of the hypoblast contributes to the formation of the coleorhiza and the remainder dries up, sticking to the end of the coleorhiza. 2. The maize embryo possesses dorsiventrality and cotyledon dimorphism. During early proembryo stage, the dorsiventrality appears in the proper of the embryo. On the ventral side, the cells are small with dense cytoplasm and few vacuoles. On the dorsal side, the cells are larger with lower cytoplasmic density and have more vacuoles. During later proembryo stage, the proper develops into two parts: the ventrum and the dorsum. The ventrum rises up from the center of the ventral side. The dorsum is composed of the marginal area of the ventral side and the whole dorsal side of the proper. During young embryo development, the ventrum differentiates into the coleoptile, apical meristem, hypocotyl, radicle and the main part of the coleorhiza. What is more important, the emergence of coleoptile primordium and radicular initials occur at the axis of the proper, then the coleoptile primordium expands from its two ends toward left and right to form a ring, and the endogenous radicular initials expand in all directions to form a conical radicular tip. All these morphogenetic activities of the ventrum follow a bilateral symmetrical pattern. The dorsum forms the scutellum primordium. Then the scutellum primordium expands rapidly toward the left, right, front and back, while thickening itself, so as to make all components originating from the ventrum become hidden in the longitudinal groove of the scutellum. Lastly, the left and right lateral scales emerge from the edges of the longitudinal groove and expand toward the central line of the axis. As a consequence, morphologically, the bilateral symmetry of the ventral side of the embryo is revealed entirely. Morphogenetically, the coleoptile primordium and apical meristem in maize are similar to the coleoptile (apical cotyledon) and apex formation of the rice embryo, so the coleoptile of the maize embryo can also be considered as an apical cotyledon. The scutellum is a lateral cotyledon. These dimorphic cotyledons of the maize embryo originate from the dorsiventrality of the proper. 3. The true morphological structure of the maize embryo is recognized and its developmental stages are established. A maize embryo is a hypocotyl, in which the apical part is the shoot apex (or plumule) with the coleoptile, the central part consists mainly of the hypocotyl with a lateral cotyledon (scutellum), and the basal part is the radicle with coleorhiza. The left and right lateral scales derived from the scutellum overlap at the ventral side, leaving only two little pores at both ends of the seam from which the coleoptile and coleorhiza can be seen. The four sequential stages of maize embryonic development are as follows: (1) proembryo stage. This stage covers a period from zygotic cell division to the appearance of the dorsum and ventrum. (2) ventrum rapid differentiation stage. (3) scutellum rapid expansion stage. (4) lateral scale development stage (or embryonic envelope formation stage). 4. To obtain a median longitudinal section perpendicular to the ventral surface is crucial for recognizing the genuine morphological structure of the maize embryo.
冯九焕 徐雪宾 刘向东 章崇玲 梁秀兰 吴万春
（1.华南农业大学植物分子育种研究中心，广州510642; 2. 华南农业大学生命科学学院，广州510642；
摘要： 运用扫描电镜与半薄切片技术,观察了玉米( Zea mays L.)的胚发育过程,得到以下认识: 第一、关于原胚.玉米合子细胞分裂形成的原胚分为胚柄、胚基与胚体三部分.胚柄短小,寿命短暂.胚基具有生长带,纵向伸长长度大,胚基的上部参与形成胚根鞘,其余部分干缩后附在胚根鞘末端.
通讯作者。Tel: 020-85280205; Fax: 020-85280212; E-mail: yglu @ scau.edu.cn.
The genomic composition of T911289, a wheat (Triticum aestivum L.) maintainer of K_CMS, was examined by several methods, such as genomic in situ hybridization (GISH), biochemical marking, and DNA molecular marking. The results got by GISH and PCR amplification of dispersed rye-specific repetitive DNA sequence suggested that the alien chromatin in T911289 derived from rye. Specifically PCR amplification of the rye-specific microsatellite primers (SCM9) and seed storage protein analysis indicated that the alien chromatin in T911289 had developed from the short arm of 1R chromosome of rye (1RS). PCR amplification by using microsatellite primers locating on 1BS and seed storage protein analysis also revealed that T911289 had lost the arm of 1BS or a small distal segment of it. We conclude that T911289 is a heterogeneous population which displays two distinct different types of translocation, i.e. the Robertsonian translocation and small segment translocation. The Robertsonian translocation type observed in our study is different from the 1BL/1RS translocation which is widely used in wheat production; it may be a novel and complex translocation form. Though the linkage between the desirable agronomic traits and the deleterious genes expressed as sticky dough has not got broken in T911289, the recovery of small segment translocation will still benefit the genetic study of wheat and rye. K型小麦细胞质雄性不育保持系T911289中外源遗传物质的初步鉴定
刘保申 李大勇 张学勇 高庆荣 孙兰珍 孙其信 董树亭
摘要： 利用荧光基因组原位杂交(GISH)、生化标记和DNA分子标记技术对普通小麦(Triticum aestivum L.)K型细胞质雄性不育保持系T911289的染色体组成进行了鉴定与分析。GISH鉴定和黑麦特异散布重复序列的检测结果表明，T911289的外源遗传物质来源于黑麦，黑麦1RS上的微卫星引物SCM9扩增结果和醇溶蛋白酸性聚丙烯酰胺凝胶电泳(A—PAGE)分析、低分子量谷蛋白的sDS_PAGE分析均表明，T911289所含的黑麦遗传物质来源于1RS；A-PAGE和SDS-PAGE分析及小麦1BS上的微卫星引物的扩增结果则表明，‘1911289缺少1BS染色体臂或1BS末端片段。GISH鉴定结果还表明，‘1911289中有罗泊逊易位和小片段易位两种类型的杂交信号，说明T911289是一个异质群体，但其罗泊逊易位又不同于生产上大面积应用的1BL／1RS易位，它可能是一种新的复杂易位形式。虽然T911289的小片段易位未能打破优异农艺性状与劣质蛋白基因的连锁，但这种小片段易位的获得将有利于小麦和黑麦的遗传研究，这种种质材料在育种上的应用价值也应优于罗泊逊易位。
The identification of germplasm is an important step for effective utilization of the available germplasm. In previous studies, isoenzyme, RAPD and SSR techniques had been used to conduct the genetic identification of watermelon (Citrullus lanatus (Thunb.) Mansf.）, but their effectiveness was limited due to the extremely narrow genetic background among watermelon genotypes. In this research, amplified fragment length polymorphism (AFLP), which was reported as a reliable technique with high efficiency in detecting polymorphism, was used to conduct genetic analysis and variety identification of thirty genotypes of watermelon core collection that represent a wide range of breeding and commercially available germplasm. As a result, a DNA fingerprint based on 15 bands amplified with four primer combinations was developed. In this fingerprint, each genotype has its unique fingerprint pattern and can be distinguished from each other. Furthermore, in order to facilitate the utilization of AFLP marker in practice, one specific AFLP band of genotype “PI296341" coming from fragment amplified by primer combination E-AT/M-CAT was successfully converted into a sequence characterized amplified region (SCAR) marker.
车克鹏 许 勇 梁春阳 宫国义 翁曼丽 张海英 金德敏 王 斌
（1. 中国科学院遗传与发育生物学研究所，北京100101；2. 北京市农林科学院蔬菜研究中心，北京100089）
摘要： 西瓜 (Citrulluslanatus (Thunb .)Mansf.)种质资源的鉴定与评价是对其有效利用的基础。以往的研究表明 ,西瓜是一种遗传资源特别狭窄的作物 ,在用同工酶、RAPD及SSR技术对西瓜种质资源进行鉴定时 ,发现很难将品种完全区分开来。本研究利用高效可靠的AFLP技术 ,对 30个西瓜核心种质材料进行了遗传分析 ,最终建立了这30个材料的DNA指纹图谱。在该图谱中 ,每个材料均有其独特的“指纹” ,材料之间可以相互区分开来。为了进一步利用AFLP分子标记 ,将重要抗病种质材料“PI2 96 341”的AFLP特异带转化成了生产上可以直接利用的SCAR标记
通讯作者。E-mail: bwang @ genetics.ac.cn。
Phosphorus is one of the three essential macroelements for plant growth. Plants respond to phosphorus starvation through adaptive mechanisms in volved in morphological, biochemical and molecular changes. To investigate the molecular background of the adaptive mechanisms, the suppression subtrac tive hybridization (SSH) method was used to construct a rice phosphorus-starvati on (Pi-starvation) induced cDNA library. Through screening of the cDNA library and sequencing of the enriched cDNAs, 18 known genes and 47 novel genes were identified. The known genes are involved in different metabolic processes, including phosphate uptake and transport, signal transduction, protein synthesis and de gradation, carbon metabolism and stress response. Northern analysis was perform ed to detect the expression patterns of some known genes and novel genes under different phosphorus levels. Different expression patterns of the selected genes were identified, which suggests that genes involved in different pathways may have different responses to Pi-starvation.
夏 铭 王首锋 王小兵 李海波 吴平
摘要： 磷素是植物生长所必需的重要元素.在缺磷环境中,植物能够调节自身的形态、生理生化和基因表达水平来适应环境的变化.为研究水稻(Oryza sativa L.)耐低磷胁迫的分子机理,采用抑制性扣除杂交技术(SSH)构建磷饥饿诱导的水稻根系扣除cDNA文库.通过文库筛选和测序获得18个已知基因和47个功能未知基因.这些基因参与了不同的代谢过程,包括磷吸收和转运、信号传导、蛋白质合成和降解、碳水化合物代谢和胁迫反应.Northern杂交结果表明,在磷饥饿胁迫下这些基因呈现不同的表达模式,并且不同代谢过程中的基因对磷饥饿有着不同的反应.
通讯作者。Fax: 0571-86971323; E-mail: <docpwu @ cls.zju.edu.cn>。
The random amplified polymorphic DNA (RAPD) technique was used to amplify DNA fragment, aiming at finding markers linked to the sex determination in Ginkgo biloba L. One thousand and two hundred random decamers had been screened. Of the 8 372 RAPD bands, only a 682 bp RAPD marker generated by a primer (S1478) of random decamer sequence, named S1478-682, was found to be associated with the male plants. This marker was present in all male plants and absent in all female plants. Ginkgo trees collected in both Beijing and Shenyang, China were tested using primer S1478. Positive results were obtained, suggesting S1478-682 could be utilized as a reliable RAPD marker to detect the sexuality of Ginkgo.
姜 凌 尤瑞麟 李懋学 时 晨
摘要：利用RAPD技术寻找银杏(Ginkgo biloba L.)中与性别相关的分子标记.筛选了1 200个10 bp的随机引物,产生了8 372个RAPD条带.只有S1478产生一条大小为682 bp、雄性特异的分子标记,该分子标记被命名为S1478-682,出现在所有雄性植株中,而所有雌性植株都不具有该分子标记.通过在北京和沈阳种植的银杏植株的RAPD推广验证,说明该分子标记可以用来检测银杏植株的性别.
通讯作者。Tel:010-62751845;Fax: 010-62751526; E-mail: rlyou @ pku.edu.cn。
Plasmid pVK1001 which carried the gfp gene of GFPmut2, a mutant of GFP,was introduced into Azospirillum brasilense Yu62 by electroporation. Maize seedlings were inoculated with the GFP-labelled bacteria and grown gnotobiotically in flask with semi-solid agar medium. Observations were performed with confocal laser scanning microscopy (CLSM) and electron microscopy, respectively, at 8 d and 12 d after inoculation. Confocal laser scanning microscopy showed that A. brasilense Yu62 could penetrate into the cortex tissue, colonizing in the intercellular spaces of the parenchyma cells of the cortex tissue. Transmission and scanning electron microscopy (TEM) showed that the majority of the bacteria colonized on the root surface and only a minority of them resided in the root interior.
刘 元 陈三凤 李季伦
摘要： 将GFPmut2质粒中的gfp基因 (编码绿色荧光蛋白)克隆到载体pVK100中,构建成重组质粒pVK1001.将pVK1001通过电转化方法导入到联合固氮菌巴西固氮螺菌Yu62中,获得GFP标记的巴西固氮螺菌Yu62菌株.用标记菌株接种限菌培养条件下生长的玉米(农大3318)幼苗,在接种后8 d、12 d,用激光共聚焦扫描显微镜进行观测,结果表明巴西固氮螺菌Yu62菌株能定植于玉米根部皮层的薄壁细胞间隙.用扫描电镜和超薄切片电镜观察表明,大多数细菌主要定植于根表,少数菌可进入玉米根组织内.
通讯作者。Tel: 010-62892715; E-mail: chensf @ maul.cau.edu.cn。
Twelve steroids, including eight ergostane-type sterols and four mono-glucosides of ergostane-type sterols, were isolated from the ethyl acetate soluble fraction of the fungus Pleurotus ostreatus (Jacq.:Fr.) Kummer. A new compound 3-O-β-D-glucopyranosyl-22 E, 24R-ergosta-7, 22-diene-5α, 6β, 9α-triol, was structurally elucidated by spectroscopic and chemical methods. The eleven known compounds were 22E, 24R-ergosta-7, 22-diene-3β, 5α, 6β, 9α-tetraol; 3-O--D-glucopyranosyl-22E, 24R-ergosta-7, 22-diene-5α, 6β-diol; 22E, 24R-ergosta-7, 22-diene-3β, 5α, 6β-triol; 22E, 24R-ergosta-5, 7, 22-trien-3β-ol 3-O-β-D-glucopyranoside; ergosterol; 22E, 24R-ergosta-7, 22-dien-3β-ol 3-O-β-D-glucopyranoside; 22E, 24R-ergosta-7, 22-diene-3β-ol; 22E, 24R-ergosta-4, 6, 8, 22-tetraen-3-one; 22E, 24R-ergosta-7, 22-diene-3β, 5α, 6α-triol; ergosterol peroxide; 22E, 24R-ergosta-7, 22_diene-3β, 5α-diol-6-one.
占扎君 王 莹 杨升平 岳建民
摘要：从口蘑科真菌北风菌(Pleurotus ostreatus (Jacq．：Fr．)Kummer)的乙酸乙酯部分分离鉴定了12个甾体类化合物，其中包括8个麦角甾醇类甾体及4个甾体苷。利用现代波谱技术及化学方法，鉴定了一个新化合物3-O-β-D-glucopyranosyl-22E, 24R-ergosta-7, 22-diene-5a, 613, 9α-triolo 其余11个已知化合物分别为22E,24R-ergosta-7, 22-di-ene-3β, 5a, 6β3, 9α-tetraol, 3-O-β-D-glueopyranosyl-22E, 24R-ergosta-7, 22-diene-5α, 6β-diol、22E, 24R-ergosta-7, 22-di-ene-3β, 5α, 6β-triol, 22E, 24R-ergosta-5, 7, 22-tfien-3β-ol 3-O-β-D-glueopyranoside, ergosterol, 22E, 24R-ergosta-7, 22-dien-3β-ol 3-O-β -D-glueopyranoside, 22E, 24R-ergosta-7, 22-diene-3β-ol、 22E, 24R-ergosta-4, 6, 8, 22-tetraen-3-one、22E, 24R-ergosta-7, 22-diene-313, 5ct, 6a-tfiol, ergosterol peroxide 和 22E, 24R-ergosta-7, 22-diene-3β, 5α-diol-6-oneo (关键词：北风菌,口蘑科,甾体,甾体苷；-O-β-D-glueopyranosy1-22E, 24R-ergosta-7,22-diene-5a,6β,9a-triol
通讯作者。E-mail: jmyue @ mail.shcnc.ac.cn。
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