The mitogen-activated protein (MAP) kinase cascades are composed of mitogen-activated protein kinases （MAPKs）, mitogen-activated protein kinase kinases （MAPKKs） and mitogen-activated protein kinase kinase kinases （MAPKKKs）, they transfer signals through phosphorylations of MAPKKK→MAPKK→MAPK. MAP kinases include a large family of serine/threonine protein kinases which are structurally conserved in eukaryotes. MAP kinase cascades play essential roles in signal transductions of extracellular signals to intracellular targets in eukaryotes. Some MAPKs, MAPKKs and MAPKKKs have been isolated from the higher plants, they mediate the signal transductions involved in plant responses to hormones, cell proliferation and differentiation, and environmental stresses.
刘强1,2 张贵友1 SHINOZAKI Kazuo 3
Protoplasts of Page tangelo (Citrus reticulata Blanco×C. paradisi Macf.) cell suspension culture were electrically fused with mesophyll protoplasts isolated from trifoliate orange (Poncirus trifoliata (L.) Raf.). More than 150 plantlets regenerated after 4-5 months of culture. The regenerated plants were trifoliate with well developed root systems. Root-tip chromosome counting of more than 20 randomly selected plants revealed that they were all tetraploids (2n=4x=36). RAPD analysis of 7 randomly selected plants verified their hybridity. Inoculation of citrus Phytophthora parasitica Dastar toxin on leaves of somatic hybrids and both parental genotypes showed that Page tangelo was moderately susceptible, and trifoliate orange was highly resistant while the somatic hybrids were resistant. The potential of this somatic hybrid as rootstock is also discussed.
郭文武 周长河 伊华林 邓秀新*
摘要：Page橘柚 (Citrusreticulata Blanco×C .paradisi Macf.)胚性细胞悬浮系原生质体与枳 (Poncirustrifoliata (L .)Raf.)叶肉原生质体经电场诱导融合 ,4～ 5个月再生 15 0余棵小植株。再生植株根系发达 ,叶片具三叶特征。随机检查 2 0余株再生苗根尖染色体数目 ,表明都为四倍体 (2n =4x =36 )。随机取 7株进行RAPD分析 ,表明被检测植株都为杂种。用引起脚腐病的寄生疫霉菌 (PhytophthoraparasiticaDastar)毒素接种体细胞杂种及双亲叶片 ,结果表明 ,Page橘柚中度感病 ,枳高抗 ,体细胞杂种为抗病类型。
The net photosynthetic rate (Pn), the apparent quantum yield (AQY), the photochemical efficiency of PSⅡ(Fv/Fm), the quantum yield of PSⅡ electron transport (Ф PSⅡ) and the coefficient of photochemical quenching (qp) were decreased in rice (Oryza sativa L.) leaves under high nocturnal temperature (42±1) ℃ stress, but the relative reduction state of PSⅡ (1-qp) was increased. With the increment of stress time, the chlorophyll content and the binding degree of chlorophyll-protein complex declined gradually, the O2(superoxide radical) production rate and H2O2 content in leaves elevated. The activities of superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) increased for 2-3 days under high temperature stress yet decreased afterwards. The results showed that the photosynthetic efficiency declined in rice leaves under high nocturnal temperature stress with concurrent emergence of the symptom of oxidative damage. The gradual increase of the SOD, POD and CAT activities, as well as that of the ratio of Pr (photo-respiration rate) to Pr+Pn and non-photochemical quenching of chlorophyll fluorescence (NPQ) indicates that those mechanisms related to the changes of these parameters may play an important role in protecting rice leaves from oxidative damage under high nocturnal temperature stress.
夜间高温 (42± 1)℃胁迫后水稻 (Oryzasativa L .)叶片的净光合速率 (Pn)下降 ,表观量子效率 (AQY)、PSⅡ原初光能转化效率 (Fv/Fm)、PSⅡ线性电子传递的量子效率 (ФPSⅡ)、光化学猝灭系数 (qp)下降 ,PSⅡ的相对还原状态 (1-qp)升高。随着夜间高温胁迫时间的延长 ,叶绿素_蛋白复合体的结合度和叶绿素含量逐渐降低 ,叶片中O-·2的生成速率和H2 O2 的含量增加 ;清除活性氧的酶类超氧物歧化酶 (SOD)、过氧化物酶 (POD)和过氧化氢酶 (CAT)在2 - 3d内的夜间高温胁迫下活性升高 ,随着胁迫时间的增加则活性下降。结果表明 :夜间高温胁迫引起水稻叶片光合效率下降 ,出现氧化伤害症状 ;而SOD、POD和CAT活性升高 ,光呼吸速率 (Pr)与Pr+Pn 的比值、叶绿素荧光非光化学猝灭 (NPQ)逐渐增加对水稻叶片是一种保护作用。
The increasement of the operation (ρValt) and the capacity of the alternative pathway (Valt) in tobacco (Nicotiana rustica L. cv. Gansu Yellow Flower) callus that treated by low temperature (4 ℃) were observed. The two electron transport pathways were changed alternatively, the cyanide-resistant respiration was the main respiratory pathway from the 2nd day to the 5th day, while the cytochrome pathway was the main pathway consistently in control callus. Cycloheximide and chloramphenicol treatment which inhibited protein synthesis resulted in dramatic and partly inhibition of Valt induced by low temperature respectively. This further proved above result that low temperature induced Valt. The same result was found when two inhibitors were used together. The analysis of heat production of callus indicated that the operation of the alternative pathway could be regulated to improve the micro-environmental temperature of tobacco callus to adapt to low temperature. The possible participation of the alternative pathway in the response to low temperature and its physiological roles were discussed.
周功克1 李红玉2 文江祁2 孔英珍1 梁厚果2*
（1. 兰州大学干旱农业生态国家重点实验室，兰州730000；2. 兰州大学生命科学学院，兰州730000)
摘要：继代培养的Nｉｃｏｔｉｏｎｏ ｒｕｓｉｔｏａ Ｌ．ｃｖ．Ｇａｎｓｕ Ｙｅｌｌｏｗ Ｆｌｏｗｅｒ）愈伤组织经低温（４℃）处理后，交替途径容量９Ｖａｌｔ）和实际运行量（ρＶａｈ）均被显著地诱导，同时其电子传递途径也在发生交替变化。低温胁迫开始时以细胞色素途径为主，至第２天时则以交替途径为主，并一直持续到第５天，以后则又转为经细胞色素途径为主，而对照却始终以细胞色素途径为主。经放红菌酮或氯霉素处理后，受低温诱导的Valt分别被显著或部分抑制，进一步表明低温诱导了Valt。用两种抑制剂共同处理也得到相同的结论。愈伤组织产热分析表明，低温胁迫下愈伤组织可调节ρVakt以提高其微环境的温度来适应低温。讨论了抗氰呼吸参与低温反应的可能生理作用。
Nostoc flagelliforme Born. et Flah is highly adapted to drought stress, cold and light stresses, and suitable for growing in the unfavorable areas. This paper presents the results of the analysis of the membrane (mainly thylakoid membrane) lipids from N. flagelliforme in order to investigate the relationship between membrane lipid composition and stress resistance to this cyanobacteria. The membrane lipids are composed of monogalactosyl diacylglycerol (MGDG), digalactosyl diacylglycerol (DGDG), sulfoquinovosyl diacylglycerol (SQDG) and phosphatidylglycerol (PG). The major fatty acids in these lipids are palmitic (16∶0), palmitoleic (16∶1), stearic (18∶0), oleic (18∶1), linoleic (18∶2) and linolenic (18∶3) acids. In N. flagelliforme, polyunsaturated fatty acids account for 73% of the total fatty acids, much higher than that of the other cyanobacteria reported so far. Among which 16∶1 and 18∶3 are as high as 28.9% and 34.3% respectively. The high resistance of N. flagelliforme to abnormal conditions may be associated with the extent of unsaturation of fatty acids. In addition, the wild N. flagelliforme treated with water for 30 min and cultured for 24 h and the lipid and fatty acid composition were found to be not affected by water-absorption.
王梅 钟泽璞* 王可玢 李良璧 匡廷云
摘要：采用改进的Allen’s的绿胶系统 ,首次对陆生蓝藻发菜 (Nostocflagelliforme Born .etFlah .)类囊体膜色素蛋白复合物进行了分离 ,共分离出了 11条绿色的色素蛋白复合物条带 ,两条浅黄色的条带。其中 7条绿色条带属于PSⅠ组分 ,4条绿色条带属于PSⅡ组分 ,1条浅黄色条带经光谱分析初步认定为类胡萝卜素蛋白复合物 ,而另一条浅黄色条带为游离色素。
Four C4 pathway enzymes of “Heinong 41" leaves of soybeans (Glycine max （L） Merr.) were assayed in five developmental stages. Pn (the net photosynthetic rate) and the chlorophyll fluorescence parameters (Fv/Fo, qP, qN and ΦPSⅡ) were also measured. The results indicated that the activities of C4 enzymes, Pn, the chlorophyll fluorescence parameters and the ratio of PEPCase (PEP carboxylase)/RuBPCase (ribulose-1,5-biphosphate) shared the same changing trait during the whole developmental stages of “Heinong 41". Correlation analysis showed that the activities of C4 enzymes, Pn, the chlorophyll fluorescence parameters were positively correlated with the ratio of PEPCase/RuBPCase. All these suggested that the degree of C4 pathway expression in “Heinong 41" leaves were positively correlated with its Pn; the photochemical function of PSⅡ was adjusted to meet the specific energy requirements for the operation of C4 pathway with the improvement of C4 expression.
李卫华1 卢庆陶1 郝乃斌1* 戈巧英1 张其德1 蒋高明1 杜维广2 匡廷云1
（1. 中国科学院植物研究所，北京100093；2. 黑龙江省农业科学院大豆研究所，哈尔滨150086）
Cytotoxin agents dactinomycin, cycloheximide and colchicine induced apoptosis processes in the meristematic cells of maize (Zea mays L.) roots were detected respectively at the chromosome, nuclear, and DNA levels simultaneously using DNA Laddering, DNA Gel Blot and chromosome spreading-based in situ end labelling. Non-random degradation of nuclear DNA (DNA Ladder) and characteristic morphological changes of chromatin/chromosome and nucleus such as condensation paralleled with those seen in apoptotic processes in animal cells, suggesting that apoptotic pathway was activated by the tested agents in maize roots in vivo.
宁顺斌 宋运淳* 王玲 刘立华
摘要：同时应用DNALaddering、DNAGelBlot以及基于染色体涂片的原位末端标记技术 ,从染色体、细胞核和DNA不同水平对细胞毒素类药物放线菌D、放线菌酮和秋水仙碱诱导的玉米 (ZeamaysL .)根尖分生组织细胞死亡作了检测。结果表明 :同动物中一样 ,这些药物诱导的玉米根尖分生组织细胞死亡也具有DNALadder、染色质和细胞核浓缩等典型的凋亡特征 ,说明这些细胞毒素类药物能够诱导植物体内的细胞凋亡。
关键词： 细胞凋亡；植物；细胞毒素类药物；原位末端标记；DNA Ladder
The effect of phospholipase C (PLC) signaling pathway on lily (Lilium davidii Duch.) pollen tube elongation was examined by means of microinjection. Pollen tube elongation was inhibited by microinjecting antibodies against animal PLCβ1-3 or inositol-1,4,5-triphosphate receptor （IP3R2, 3）, but was not affected by antibodies against animal PLCβ4 or IP3R1. Pollen tube elongation was also stimulated significantly by microinjecting IP3. The results suggest that PLC-IP3 signaling pathway might present in pollen system and be involved in pollen tube growth.
王昕 崔素娟 马力耕 孙大业*
摘要：以百合 (Liliumdavidii Duch .)花粉为材料 ,通过显微注射肌醇磷脂信使系统中重要组成成分或其抗体 ,研究该信使系统对花粉管伸长的影响。发现显微注射动物来源磷脂酶C(PLC) β1-3 抗体显著抑制花粉管的伸长生长 ,而注射PLCβ4 抗体对花粉管伸长无影响 ;显微注射三磷酸肌醇 (IP3 ) ,可显著促进花粉管伸长生长 ;显微注射动物来源的IP3 R2 、IP3 R3 抗体可显著抑制花粉管伸长生长 ,而注射IP3 R1抗体对花粉管伸长无影响。上述结果表明肌醇磷脂信号系统可能参与花粉管伸长生长过程。
The Raphanus sativus L. antifungal protein 1 (Rs-AFP1) gene was isolated by polymerase chain reaction (PCR). The complete open reading frame and the fragment encoding the putative mature protein were inserted into the prokaryotic expression vector pET-32b(+), respectively. Subsequent expression showed that the Rs-AFP1 was produced in E. coli as a 27 kD fusion protein only when the N-terminal signal peptide was removed. After treatment with thrombin to remove part of the N-terminal His.tag sequence, the bacterially expressed Rs-AFP1 was used for fungal growth inhibition assay which was conducted on Verticillium dahliae Kleb., a soil-born fungus causing the cotton wilt disease. Results showed that, in the liquid medium, the Rs-AFP1 fusion protein at a concentration of 0.3 g/L clearly inhibited the growth of V. dahliae and the germination of spores. Thus the bacterially expressed fusion protein had the antifungal activity against V. dahliae.
周向军1 王仑山2 林芝萍1 贾军伟1 卢山1 储昭庆1 陈晓亚1*
摘要：利用聚合酶链式反应（ＰＣＲ）获得了萝卜（Ｒａｑｈａｎｕｓ ｓａｔｉｖｕｓ Ｌ．）抗真菌蛋白１（Ｒｓ－ＡＦＰ１）基因编码区核苷酸序列。将整个阅读框架片段和云除了Ｎ－端信号肽序列的片段分别装入原核表达载体ｐＥＴ－３２ｇ（＋）中，在大肠杆菌中表达，发现带有信号 的Ｒｓ－ＡＦＰ１不能在大肠杆菌中表达，而当这一序列去除后，表达出约２７ｋＤ的Ｒｓ－ＡＦＰ１的融合蛋白。用凝血酶处理融合蛋白以云除Ｎ－端Ｈｉｓ．ｔag的部分序列，然后用处理后的融合蛋白进行了抑制真菌生长的实验。结果表明，在加入0.3g/L的Rs-AFP1的融合蛋白的培养液中，大丽轮枝菌（Vertivillium dahloae Kleb.）的生
A complete set of telotrisomics, developed from rice (Oryza sativa L.ssp. indica) variety, “Zhongxian 3037", are great valuable genetic stocks for rice genetic studies. The authors have established a direct and simple mapping approach to assign various kinds of rice DNA sequences to definite chromosome arms according to their dosage effects. By use of the rice telotrisomics, three types of rice DNA clones reluctant to linkage analysis were assigned to specific chromosome arms, respectively. Among them, RG684, a clone showing sever distorted segregation, was assigned to chromosome arm 10L; two genomic respective copies of a multiple-copy clone, G1073, were assigned to 1L; and RCS1, a cereal centromeric repetitive clone, was assigned to 4L.
程祝宽1,2 颜辉煌1 于恒秀2 钱前1,3 裔传灯2 顾铭洪2 朱立煌1*
（1. 中国科学院遗传研究所，北京100101；2. 扬州大学农学院农学系，扬州225009；3. 中国水稻研究所，杭州310006）
摘要：以水稻 (Oryzasativa L .ssp .indica)“中籼 30 37”成套端三体为材料 ,通过制备不同染色体两个端三体的等量DNA膜 ,并与待定位的分子标记或DNA序列杂交 ,成功地将严重偏分离的零等位标记RG6 84、多拷贝标记G10 73的不同片段 ,以及着丝点相关重复序列RCS1定位到相应的染色体臂上。说明在水稻中 ,不同染色体臂的端三体可以弥补连锁定位方法的一些不足。
Cauliflower curd is composed structurally of a number of shortened shoots on which there are tremendous amount of naked apical meristems. The authors isolated four pure lines of different curd types from cauliflower (Brassica oleracea L. var. botrytis) accessions: smooth curd (curd-s), haired curd (curd-h), crystal curd (curd-c) and thorned curd (curd-t). BobCAL genes was cloned separately from each pure line by PCR amplification and a termination mutation from AAG to TAG was found in the fifth exon of all four sequences, whereas DNA sequences of 638 bp upstream of the stop codons were identical, demonstrating that BobCAL is not the sole regulator of flower buds and cauline on curd. After crosses of cauliflower with cabbage (B.oleracea var. capitata) or broccoli (B.oleracea var. italica), the resultant F1 plants failed to form curd and had the diversity of inflorescence which implicates the relatedness of CAL to other regulators. One homologous gene of CAL, BcpCAL was isolated from Chinese cabbage (B.campestris L. ssp. pekinensis) and it was found not to have the termination mutation in the fifth exon, and thus the structure of BcpCAL gene was analyzed. The results offer molecular and genetic evidence for the study of biological function of CAL and morphological genetics of curd.
李小方 薛万新 孙玉东 沈瑞娟 刘平林 汤章城 何玉科*
摘要：从花椰菜变种 (Brassicaoleracea L .var.botrytis)中分离出 4种花球类型的纯合株系 :光滑型花球curd-s、毛状物花球curd-h、颗粒状花球curd-c和刺状物花球curd-t。用PCR方法分别扩增各株系的BobCAL基因并分析其中的部分序列 ,发现 4种株系的CAL基因在第五个外显子中都有AAG向TAG的终止突变 ,而且该终止密码子上游 6 38bp的DNA序列完全一致 ,说明花蕾、茎生叶等花球附生物的形态发生不是BobCAL的单一调节作用。花椰菜与结球甘蓝 (B .oleracea var.capitata)和青花菜 (B .oleracea var.italica)杂交后其F1代植株都失去了原有花球的形态特征 ,表现出花序的多态性 ,从而显示了不同变种CAL相互作用的形态学差异。从大白菜中分离出CAL的同源基因 ,发现BcpCAL基因没有发生终止突变。分析了从大白菜 (B .campestris L .ssp .pekinensis)中分离出的BcpCAL的基因结构特征以及与其他基因间的同源性。为了解CAL基因的生物学功能 ,研究花球形态发生的机制提供了分子和遗传证据。
关键词： 花椰菜；CAL 基因；花球；大白菜
The plasmid pRSSGNA1 carried a snowdrop lectin gene (Galanthus nivalis agglutinin, GNA) under the drive of RSs-1 promoter, were successfully transferred into three tomato (Lycopersicon esculentum Mill.) cultivars, “C8”，“A39” and “A53” via Agrobacterium-mediated transformation. Regenerated plantlets were obtained from cotyledons after preculture, shoot inducing culture and root inducing culture. Transgenic tomato plants were confirmed by the kanamycin-resistant experiment, PCR analysis and Southern blot. The preliminary results from bioassay demonstrated significant resistance of the transgenic plants to aphid (Myzus persicae Sulzer) larvae. The inheritance of selective marker gene (NPTⅡ) in 3 transgenic tomato plants is in the model of the simple Mendel's fashion in progenies of the selfing generation.
吴昌银1 叶志彪1* 李汉霞1 唐克轩2
（1. 华中农业大学作物遗传改良国家重点实验室，武汉430070；2. 复旦大学遗传研究所，上海200433）
摘要：将含有雪花莲外源凝集素基因 (GNA)的质粒pRSSGNA1通过冻融法转化到根癌土壤杆菌 (Agrobacteriumtumefaciens (SmithetTownsend)Conn)菌株LBA44 0 4中。采用叶盘法转化番茄 (Lycopersiconesculentum Mill.)栽培品种“C8”、“A3 9”和“A53 ” ,获得了含GNA基因的 43株转化植株。通过卡那霉素抗性鉴定、NPTⅡ基因PCR和GNA基因Southernblot分析表明 ,GNA基因已整合到番茄的基因组中。初步抗蚜虫 (MyzuspersicaeSulzer)试验证明 ,转基因番茄有一定的抗蚜虫效果。 3株转基因植株的NPTⅡ基因在自交子代中的分离比例符合 3∶1孟德尔遗传规律。
A maize (Zea mays L.) genome-specific repeated DNA sequence (clone MR64) has been transferred into one DH line of wheat through wheat (Triticum persicum Vav. ex Zhuk.) and maize cross. In the present study by RFLP analysis the authors proved that this DNA sequence could stably transmit into DH3 plants, the next generation derived from DH2 self-crossing. A similarity search in all DNA databases using BLASTN program showed that the DNA sequence of MR64 had as high as 93% identity to PREM-2 and 79% to Opie-2 in nucleotides. Both PREM-2 and Opie-2 are known as retrotransposons in maize genome, suggesting that MR64 likely is the partial sequence of a maize retrotransposon. Therefore, the results indicate that some retrotransposon might involve the DNA introgression from maize to wheat genome through wide fertilization. Stable inheritance of this maize genome-specific retrotransposon-like DNA in the wheat genome opens up the possibility of using retrotransposon as a new tool for gene tagging, function analysis, and insertional mutagenesis in wheat genome.
陈纯贤1,2 孙敬三2 朱立煌1*
（1. 中国科学院遗传研究所，北京100101；2. 中国科学院植物研究所，北京100093）
摘要：作者曾报道了一个玉米（Ｚｅａ ｍａｙｓ Ｌ．）基因组特异的重复序列ＤＮＡ（克隆ＭＲ６４）通过小麦（Ｔｒｉｔｉｃｕｍ ｐｅｒｓｉｃｕｍ Ｖａｖ．ｅｘ Ｚｈｕｋ．）与玉米杂交导入一个小麦加倍单倍体（ＤＨ）植株中。利用分析主宰该重复ＤＮＡ序列可以稳定地传递到小麦ＤＨ３代植株。通过Ｉｎｔｅｒｍｅｔ在ＤＮＡ数据库中进行序列相似性搜寻和同源性比较，结果显示，ＭＲ６４的ＤＮＡ序列和玉米的最近报道的两个逆转座子PREM-2和QPIE-2分别有高达93%和79%的序列同一性。因此，该玉米DNA片段很可能是一种逆转座子的一部分，这意味着有可能利用玉米的某些基因组特异的逆转座子的序列在小麦基因组中进行基因标签、功能分析和插入突变的研究。
The BmK IT4 gene was obtained from pBS-BmK IT4 by EcoRⅠ/KpnⅠdigestion and it was then cloned into pE3 intermediate vector. The resulting plasmid was named pE3-BmK IT4. The chimeric gene was transferred into the tobacco (Nicotiana tabacum L.) genome via Agrobacterium-mediated transformation. Forty-five regenerated kanamycin resistant plants were obtained, two individual lines showed strong toxicity to Manduca sexta (Linnaeus), Heliothis armigera (Hübner) and Leguminivora glycinivorella (Matsumura) by feeding experiments. Results from Southern blot indicated that BmK IT4 gene was transferred into tobacco genome. The mortality of M．sexta, H．armigera and L．glycinivorella larvae fed on transgenic plants was 95%-97%, 63%-70% and 65%-73%, respectively, and the growth of the surviving insects was remarkably retarded.
胡廷章1,2 徐云剑1 江国英1 朱玉贤1*
摘要：用酶切方法从pBS-BmKIT4 质粒中获得BmKIT4 基因片段 ,并构建CaMV 35S启动子下的BmKIT4 基因表达质粒pE3_BmKIT4 ,以根癌土壤杆菌 (Agrobacteriumtumefaciens (SmithetTownsend)Conn)介导的叶盘法转化云烟 (Nico tianatabacumL .)K32 6叶片 ,获得 45株抗卡那霉素的再生植株。用这些再生植株进行抗虫试验 ,发现有 2株植株对烟草天蛾 (Manducasexta (Linnaeus) )、棉铃虫 (Heliothisarmigera (H櫣ｂｎｅｒ) )和大豆食心虫 (Leguminivoraglycinivorella(Matsumura) )都有较强的抗性。收获这 2株烟草的自交种子后 ,对T1代幼苗进行Southern杂交分析表明 ,BmKIT4基因已转入这些烟草中 ,用这些子代植株进行抗虫试验 ,与对照株相比 ,这 2株转基因烟草对烟草天蛾的杀虫率达95 %～ 97% ,对棉铃虫的杀虫率达 6 3%～ 70 % ,对大豆食心虫的杀虫率达 6 5 %～ 73% ,并能显著抑制昆虫的蜕皮和生长发育 ,表现出强烈的抗虫作用。
关键词： BmKIT4 基因；转基因烟草；抗虫性
The ability of plants to undergo androgenesis is influenced by their genotypes, which is thus very important in anther culture response. However, little is known about the mechanism underlying genotypic effects. In this study, two barley (Hordeum vulgare L.) genotypes with different abilities undergone microspore embryyogenesis were tested. Estimation of the frequency of different pollens in anthers cultured in vitro suggested that there was no significant difference between early androgenesis of the two genotypes. Therefore, the big difference between the numbers of calli was assumed to rise after the formation of multinucleate-pollen. To find molecular markers linked to the high-frequent microspore embryogenesis, a segregated F2 population was analyzed using the bulked segregant analysis (BSA) and RAPD. Using a primer S500 (5′-TCGCCCAGTC-3′), an approximate 650 bp fragment was amplified which cosegregated with the high-frequent microspore embryogenesis.
孙海鹏 路铁刚 孙敬三*
摘要：以具有不同花粉胚胎发生能力的两个大麦 (Hordeumvulgare L .)基因型为材料 ,对离体培养花药中不同类型小孢子频率的连续统计表明 ,两基因型的早期雄核发育过程无明显区别 ,最终出愈率 (平均每个花药产生的愈伤组织块数 )的显著差异是在小孢子形成多核花粉以后产生的。推测基因型之间胚胎发生能力的差异不受小孢子脱分化启动能力控制 ,而是由其多核花粉发育成愈伤组织的能力决定的 ,可能与细胞壁的形成有关。利用BSA(bulkedsegregantanalysis)法对F2 分离群体进行分析 ,找到一个可能与高频率胚胎发生能力连锁的RAPD标记。
Random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) methods were applied to detect genetic variation of 20 bulking samples and two individually sampled populations of Oryza granulata (Nees et Arn. ex Watt.) from China (M5 from Hainan Province and M27 from Yunnan Province). For the bulking sampled populations, 20 RAPD and 12 ISSR primers generated 209 and 122 bands, of which 134 (64.11%) and 89 (72.95%) were polymorphic respectively. For population M5, 146 and 95 bands were generated with 21.48% and 34.78% being polymorphic (PPB). For population M27, 151 (PPB = 17.22%) and 94 (PPB = 19.15%) bands of RAPD and ISSR were obtained. The results indicated that the level of genetic variation of O． granulata was lower than other species detected in the genus Oryza. UPGMA cluster based on genetic similarity and principle component analyses (PCA) based on band patterns divided the populations into two groups corresponding to their sources, which revealed that the majority of genetic variation of O. granulata occurred between Hainan and Yunnan. Consequently, more populations of the species should be considered for a reasonable conservation management. Comparison between the two marker systems showed that ISSR was better than RAPD in terms of reproducibility and ability of detecting genetic polymorphism. Mantel test revealed that the goodness of fit between them was significant (r=0.917, t=12.718) when detecting genetic diversity at species level, but was poor at population level (r<0.200). The discrepancy was considered as the facts that different fragments were targeted by RAPD and ISSR, and that evolutionary process of O. granulata varied at different hierarchical levels.
钱韦 葛颂* 洪德元
摘要：用随机扩增多态DNA (RAPD)和inter_简单重复序列 (ISSR)标记对分别来自中国海南和云南 2 0个居群的疣粒野生稻 (Oryzagranulata (NeesetArn .exWatt.) )混合样品 ,以及海南 (M5 )和云南 (M2 7) 2个居群各 2 0个植株的遗传多样性进行了检测。在混合取样的居群中 ,2 0个RAPD引物和 12个ISSR引物分别扩增出 2 0 9和 12 2条带 ,多态条带比率 (PPB)分别为 6 4.11%和 72 .95 %。对于分单株取样的居群 ,在M5中 ,两种标记分别扩增出 146和 95条带 ,PPB值分别为 2 1.48%和 34 .78%。在居群M2 7中 ,则分别扩增出 15 1(PPB =17.2 2 % )和 94(PPB =19.15 % )条带。结果表明 ,中国的疣粒野生稻在物种水平的遗传多样性较低 ,其遗传变异主要存在于海南和云南两地之间 ,而在居群内的遗传多样性水平很低。基于Jaccard遗传相似性系数的UPGMA聚类和基于多态条带表型的主成分分析 (PCA)都将中国的疣粒野生稻分为来自海南和云南两类 ,两地间该物种的平均遗传相似性分别为 0 .5 96(RAPD)和 0 .5 11(ISSR) ,其间产生了相当程度的遗传分化。对比RAPD和ISSR在PCR反应中的稳定性和检测变异的能力表明 ,对于实验条件的稳定性而言ISSR优于RAPD ,且总的来说ISSR能检测到比RAPD更多的遗传变异。Mantel检测表明 ,在物种水平 ,这两种标记的分析结果有极显著的相关性（r=0.917,t=12.789）；而在居群水平，则分析结果极不一致（r<0.200）。这与它们所扩增的相应基因组片断的变异方式有关。同时，也反映了疣粒野生稻居群内和居群间存在不同的进化模式。根据以上结论，在进行中国的疣粒野生稻原位保护选址和迁地保护的取
关键词： 疣粒野生稻；RAPD；ISSR; 遗传多样性；保护生物学
Net N mineralization and nitrification were determined using the closed-top PVC tube in situ incubation method in a subalpine meadow (Saussurea iodostegia Hance+ Carex capillaris L.)→shagspine peashrub (Caragana jubata (Pall.) Poir) shrubland→ribbed birch (Betula costata Trautv.) successional sere. The ability of the three communities to supply available N was comparatively studied. The results showed that there were apparent seasonal changes in the inorganic N pools (including NH+4-N and NO-3-N) and net N mineralization and nitrification rates in the three sites. There were generally no significant differences in the inorganic N pools among sampling events. But the NH+4-N concentration in both birch (P<0.01) and meadow (P<0.01) sites was significantly higher than that in peashrub site in June 1996, and the NO-3-N concentration in peashrub site was significantly higher than that of meadow site (P<0.05) in August 1996. The annual net N mineralization and nitrification rates in peashrub site (16.01 kg·hm-2) were higher than in birch (12.05 kg·hm-2) and meadow sites (1.64 kg·hm-2). The annual net nitrification rate in peashrub site (11.37 kg·hm-2) was higher than in meadow site (10.90 kg·hm-2) and much lower than in birch site (14.36 kg·hm-2). We conclude that the ability of peashrub shrubland soil to supply available N for plant uptake and the ability to prevent available N from denitrification and leaching were higher than that of the other two sites. The leguminous peashrub might play a potential role in supplying more N, which in turn facilitate the invasion of birch saplings during the successional processes.
苏波 韩兴国 渠春梅 黄建辉
摘要：利用PVC管顶盖埋管原位培育法测定了东灵山顶亚高山草甸 (紫苞风毛菊(SaussureaiodostegiaHonce) +丝柄苔草 (CarexcapillarisL .)_箭叶锦鸡儿 (Caraganajubata (Pall.Poir.) )灌丛_硕桦林 (BetulacostataTrautv .)演替序列中土壤有机N的年度净矿化与硝化作用 ,并以之作为土壤供氮能力的指标 ,比较了锦鸡儿灌丛与硕桦林和草甸土壤的供氮能力和维持氮素的能力。结果表明 ,3个生态系统土壤无机氮库 (包括NH+ 4 N和NO-3 _N)及净N矿化与硝化速率都存在明显的季节变化 ;除 1996年 6月硕桦林 (P 0 .0 1)和草甸NH+ 4_N显著高于锦鸡儿灌丛 (P 0 .0 1) ,1996年 8月锦鸡儿灌丛NO-3 _N显著高于草甸 (P 0 .0 5 )外 ,在不同取样时期无机氮库大小在 3个生态系统之间都不存在显著差异 ;锦鸡儿灌丛每公顷的年度总矿化量 (16 .0 1kg·hm-2 )高于硕桦林 (12 .0 5kg·hm-2 )和草甸 (1.6 4kg·hm-2 ) ;净硝化量 (11.37kg·hm-2 )略高于草甸 (10 .90kg·hm-2 ) ,低于硕桦林 (14.36kg·hm-2 )。尽管锦鸡儿灌丛土壤无机氮含量 ,矿化、硝化速率并不明显高于硕桦林和草甸 ,但其总年度净矿化量最高 ,所以锦鸡儿灌丛土壤的供氮能力在 3个群落中最强。此外 ,由于锦鸡儿灌丛的总年度硝化量低于硕桦林 ,略高于草甸 ,因此 ,锦鸡儿灌丛维持无机氮素的能力也较硕桦林强。总之，锦鸡儿灌丛通过提高氮素的可利用性而推动该序列的演替展。
关键词： 净9 矿化与硝化作用；锦鸡儿灌丛；演替
The Schisandraceae is one of the most important taxa for understanding the origin and evolution of primitive angiosperms due to its basal position in the recent cladograms of the angiosperm based both on several gene sequences and on morphological characters, but phylogenetic relationships within the family are still unresolved. The sequences of nrDNA ITS region of 15 species representing four sections of Schisandra Michx., two sections of Kadsura Kaempf. ex Juss. and one outgroup, Illicium fargesii Finet et Gagnep., were used to reconstruct the phylogeny of Schisandraceae. Fourteen most parsimonious trees (Length=259, CI=0.934 and RI=0.889) were obtained from the analysis with I．fargesii as the outgroup. In the consensus tree, the genus Schisandra was found to be divided into three clades. Sect. Pleiostema formed a clade together with sect. Maximowiczia, sect. Sphaerostema was weakly supported to be the sister group of a clade comprising S. bicolor var. tuberculata and Kadsura species. In particular, S. bicolor var. tuberculata, a species of sect. Schisandra, was nested within Kadsura. It seems from this result and the morphological characters that Schisandra might not be a monophyletic group. According to the present molecular phylogeny, both elongated hypocarpium and deciduous habit originated independently at least twice in the Schisandraceae, and therefore, the value of the present morphological characters used in the classification of the family Schisandraceae should be reevaluated.
刘忠1 汪小全1* 陈之端1 林祁2 路安民1
（1. 中国科学院植物研究所系统与进化植物学开放研究实验室，北京100093；2.. 中国科学院植物研究所标本馆，北京100093）
The internal transcribed spacer (ITS) regions of nuclear ribosomal DNA from ten species of Chloranthus Swartz and two outgroup species of Sarcandra Gardn. were sequenced. The regions ranged in length from 654 to 663 bp, with ITS1 (285-290 bp) slightly longer than ITS2 (205-211 bp). Of the 669 aligned positions, 109 sites (56 in ITS1, 52 in ITS2, and one in 5.8S region) were phylogenetically informative. Only one most parsimonious tree of 252 steps was obtained in PAUP analysis when gaps were treated as the fifth state. The traditional division of Chloranthus on the basis of habit seems to be quite unnatural. Evidence from the sequence of ITS region, just as that from cytology and anatomy, strongly suggested the separation of the genus into two groups according to the characteristics of androecial organs: one contains C. angustifolius, C. nervosus, C. japonicus and C. fortunei, and the other comprises the remaining species.
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