Comparison of saponin composition and content in wild soybean (Glycine soja Sieb. and Zucc.) before and after germination

Eight wild soybean accessions with different saponin phenotypes were used to examine saponin composition and relative saponin quantity in various tissues of mature seeds and two-week-old seedlings by LC–PDA/MS/MS. Saponin composition and content were varied according to tissues and accessions. The average total saponin concentration in 1?g mature dry seeds of wild soybean was 16.08?±?3.13?μmol. In two-week-old seedlings, produced from 1?g mature seeds, it was 27.94?±?6.52?μmol. Group A saponins were highly concentrated in seed hypocotyl (4.04?±?0.71?μmol). High concentration of DDMP saponins (7.37?±?5.22?μmol) and Sg-6 saponins (2.19?±?0.59?μmol) was found in cotyledonary leaf. In seedlings, the amounts of group A and Sg-6 saponins reduced 2.3- and 1.3-folds, respectively, while DDMP?+?B?+?E saponins increased 2.5-fold than those of mature seeds. Our findings show that the group A and Sg-6 saponins in mature seeds were degraded and/or translocated by germination whereas DDMP saponins were newly synthesized.     

不同生态环境野生大豆的结构比较研究

对生长在不同生态环境的蝶形花科Ｆａｂａｃｅａｅ 大豆属Ｇｌｙｃｉｎｅ 的两个野生大豆Ｇ．ｓｏｊａ 品系进行了扫描电镜观察及比较研究．结果表明,生长在盐渍生态环境的野生大豆茎和叶体表都具有盐腺,盐腺圆球型,基部有一个小柄,着生在盐生野生大豆茎、叶表皮外切向壁的胞间层处．幼嫩的盐腺靠泌盐孔泌盐,成熟的盐腺靠整体破碎释盐．而生长在黑土地生态环境的野生大豆的茎和叶外切向壁未发现有泌盐的盐腺,其茎叶的表皮都呈现出平滑状态．因此两种不同生态环境的同科同属植物在微观结构上显示出明显差异．     

野生大豆种子雨的研究

对野生大豆种子雨的时空动态及其与外界天气情况关系的初步研究表明,在野 生大豆整个种子雨的历程中,出现３个较为明显的炸荚和种子散落的高峰,并且峰的出现 与天气晴朗（相对湿度）相关,而与气温相关性不大．野生大豆种子雨的空间分布格局主要 是野生大豆种子炸荚的自身习性（炸荚弹力）和荚本身在植株上的空间分布有关,而与风 向（风力≤７级）关系不大．     

野生大豆种子雨的研究

对野生大豆种子雨的时空动态及其与外界天气情况关系的初步研究表明,在野生大豆整个种子雨的历程中,出现3个较为明显的炸荚和种子散落的高峰,并且峰的出现与天气晴朗(相对湿度)相关,而与气温相关性不大.野生大豆种子雨的空间分布格局主要是野生大豆种子炸荚的自身习性(炸荚弹力)和荚本身在植株上的空间分布有关,而与风向(风力≤7级)关系不大.     

渤海湾津唐沿海野生大豆(Glycine soja)种群高盐碱胁迫反应

以渤海湾津唐沿海地带895份野生大豆种群在高盐碱(3%)环境下的全生育期鉴定试验,分析不同种群(株系)的生长发育动态以及对盐碱土壤生态适应性的分化。结果表明,出苗阶段存在3种耐盐碱能力类型:耐高盐碱胁迫发芽出苗;耐低盐碱胁迫发芽出苗;盐胁迫不能发芽出苗。在高盐胁迫下营养生长早期阶段种群死亡率最高,中期以后相对降低,成熟期死亡率较低。种群植株形态建成和生殖生长受到高盐胁迫的强烈抑制,表现早期严重而后期较轻。植株和产量性状的受害程度由大到小依次为:单株的地上干物质重、单株产量、种子数、百粒重和收获指数。株系间变异系数从大到小依次为:单株产量、种子数、荚数、收获指数、地上干物质重、百粒重。研究揭示该地域种群内对盐碱土壤有很高的适应性分化,存在高耐盐碱生态型。     

野大豆(Glycine soja Sieb.et Zucc.)一新变种

命名发表了野大豆一新变种,即闭花野大豆Glycine soja Sieb.et Zucc.var.cleistogama C.S.Zhu et S.X.Zhu.     

耐盐野生大豆(Glycine soja)的光周期效应

以原产山东东营的耐盐野生大豆为材料 ,观察了不同日长和不同光周期数处理对植株开花时间、数量、结荚率、鼓粒率以及生长和光合速率的影响 :(1)短日照处理促进野生大豆开花 ,其开花临界日长是 13h ;(2 )在 8h日长处理条件下野生大豆开花所需最小光周期数是 10个 ,且随着处理光周期数的增加 ,促进开花和开花后发育的效应有逐步增强的趋势 ,不足的光周期数处理可以引起成花逆转现象 ;(3)经短日处理的野生大豆植株生长减慢、光合速率降低 ,且随着日长缩短 ,生长和光合速率的下降程度增加     

关于野大豆盐腺问题的探讨

以中国3个省的盐生野大豆(Glycine soja Sieb. et Zucc.)为材料,在沙基培养、溶液培养和大田种植3种种植条件下用不同浓度的盐处理,观测了茎叶表面附着物的形态分布和腺毛的超微结构,测定了叶片腺毛分泌物中和叶片组织内部Na+和Cl-的含量变化,并对腺毛的3个细胞以及表皮细胞和叶肉细胞内的Na+相对含量变化进行了X射线微区分析.结果发现:盐生野大豆茎叶表皮上生长的附着物中只有表皮毛和腺毛,腺毛的形态类似于禾本科植物中的一些盐腺,叶片上的腺毛均生长在叶脉上;腺毛细胞内部结构具有一般盐腺的特点,如有大液泡,稠密的细胞质,大量的线粒体、叶绿体、胞间连丝以及较厚的细胞壁等.通过测定在无盐对照、盐处理和盐处理加盐腺泌盐抑制剂条件下盐生野大豆叶片腺毛分泌物中和叶片组织内部的Na+和Cl-含量,结果显示,盐生野大豆腺毛具有泌盐功能,加入泌盐抑制剂后,其泌盐作用停止;腺毛的3个细胞以及表皮细胞和叶肉细胞内的Na+在不同的盐浓度下的微区定位分析结果表明,盐生野大豆叶片的腺毛细胞有较强的积累Na+的能力.综合分析认为,盐生野生大豆的腺毛就是具有泌盐功能的盐腺,没有发现其他类型的盐腺.     

NaCl胁迫下野生和栽培大豆幼苗体内离子的再转运

采用NaCl根际处理和叶面饲喂^22Na方法,研究了野生大豆(Glycine soja)——耐盐的BB52、盐敏感的N23232和栽培大豆(Glycine max)——较耐盐的Lee68幼苗在盐胁迫及解除过程中对Na^ 、Cl^-的吸收和再转运。结果表明,在NaCl根际处理12h过程中,BB52和Lee68幼苗根对Na^ 、Cl^-吸收和向茎、叶的运输逐渐增加,10h时趋于稳定,Na^ 、Cl^-含量高低顺序是根＞茎＞叶。但N23232的Na^ 、Cl^-含量则是茎＞根＞叶。在用NaCl对根处理10h后再解除NaCl处理的0～36h内,BB52吸收的Na^ 、Cl^-较多地留于根部或转运至根茎过渡区,叶中较少。N23232吸收的Na^ 较多地转运至茎部,而Cl^-含量在幼苗各部分无差异。叶片饲喂^22Na 10h后,BB52吸收^22Na较N23232多,并较多地向根部运输。从离子再转运角度讨论了BB52的耐盐性。     

关于野大豆盐腺问题的探讨

以中国3个省的盐生野大豆(Glycine soja Sieb. et Zucc.)为材料,在沙基培养、溶液培养和大田种植3种种植条件下用不同浓度的盐处理,观测了茎叶表面附着物的形态分布和腺毛的超微结构,测定了叶片腺毛分泌物中和叶片组织内部Na^ 和Cl^-的含量变化,并对腺毛的3个细胞以及表皮细胞和叶肉细胞内的Na^ 相对含量变化进行了X射线微区分析。结果发现：盐生野大豆茎叶表皮上生长的附着物中只有表皮毛和腺毛,腺毛的形态类似于禾本科植物中的一些盐腺,叶片上的腺毛均生长在叶脉上;腺毛细胞内部结构具有一般盐腺的特点,如有大液泡,稠密的细胞质,大量的线粒体、叶绿体、胞间连丝以及较厚的细胞壁等。通过测定在无盐对照、盐处理和盐处理加盐腺泌盐抑制剂条件下盐生野大豆叶片腺毛分泌物中和叶片组织内部的Na^ 和Cl^-含量,结果显示,盐生野大豆腺毛具有泌盐功能,加入泌盐抑制剂后,其泌盐作用停止;腺毛的3个细胞以及表皮细胞和叶肉细胞内的Na^ 在不同的盐浓度下的微区定位分析结果表明,盐生野大豆叶片的腺毛细胞有较强的积累Na^ 的能力。综合分析认为,盐生野生大豆的腺毛就是具有泌盐功能的盐腺,没有发现其他类型的盐腺。     

Restriction Endonuclease Digestion of Amplification Products Generated by RAPD Technique in a Population of Glycine soja

n a population of Glycine soja L., the polymorphic loci could be hardly detected by RAPD markers, using several primers. These non-polymorphic amplification products were cleaved by some restriction endonuclease, such as Msp Ⅰ , Hinf Ⅰ , Taq Ⅰ , EcoR Ⅰ , Sal Ⅰ , Dra Ⅰ and Hae Ⅲ. After cleaving, the digested amplification products were detected on polyacrylamide gel electrophoresis with silver staining. It was found that: 1 ) some restriction endonucleases could not, and some others could effectively digest the random amplication products of the DNAs of G. soja; 2) some endonucleases could produce polymorphic DNA fragments after digestion of the non-poly-morphic products, but others could not even after digestion; 3) non-polymorphic amplification products amplified by some primers could produce polymorphic DNA fragments after digestion, while those by other primers could not. It could be concluded that the restriction endonuclease digestion of amplification products could increase significantly detectability of polymorphic DNA by RAPDs technique.     

氯化钠胁迫下野生和栽培大豆幼苗体内的多胺水平变化

以通用的较耐盐的栽培大豆Lee68品种和对盐敏感的野生大豆N23232种群为参照,研究了盐胁迫下耐盐野生大豆BB52种群幼苗体内多胺(PAs)组分、含量及多胺氧化酶(PAO)活性的变化。结果表明,盐胁迫下BB52幼苗根PAs中Put和Spm含量下降较Lee68和N23232显著,但Spd含量下降较少．BB52叶片PAs中Put含量下降,Spd上升,(Spd+Spin)／Put值增加和Put／PAs值降低幅度与耐盐性呈正相关趋势．盐胁迫下,各材料根和叶中PAO活性上升,N23232上升最明显．探讨了多胺水平与BB52耐盐性的关系。     

野生大豆种子cDNA文库的构建与分析

为了分离与鉴定野生大豆优良基因,以双高型优质野生大豆的近成熟种子为材料,采用裂解法提取了总RNA;以Oligo（dT）为引物,经SA—PMPS法分离出mRNA,反转录酶催化合成cDNA,并以cDNA第一链为模板在DNA聚合酶Ⅰ的作用下合成cDNA第二链,双链cDNA经加接头等步骤,成功构建了野生大豆cDNA文库。文库的重组率约为93．7％,PCR检测重组克隆的插入片段平均大于1000bp,测序片断大于500bp,表明构建的近成熟种子cDNA文库质量较高,为进一步进行EST测序和全长克隆打下了基础。     

The genomic relationship between Glycine max (L.) Merr. and G. soja Sieb. and Zucc. as revealed by pachytene chromosome analysis

Summary This study was conducted with the objective of determining the genomic relationship between cultivated soybean (Glycine max) and wild soybean (G. soja) of the subgenus Soja, genus Glycine. Observations on cross-ability rate, hybrid viability, meiotic chromosome pairing, and pollen fertility in F ₁ hybrids of G. max × G. soja and reciprocals elucidated that both species hybridized readily and set mature putative hybrid pods, generated vigorous F₁ plants, had a majority of sporocytes that showed 18II + 1IV chromosome association at diakinesis and metaphase I, and had a pollen fertility that ranged from 49.2% to 53.3%. A quadrivalent was often associated with the nucleolus, suggesting that one of the chromosomes involved in the interchange is a satellited chromosome. Thus, G. max and G. soja genetic stocks used in this study have been differentiated by a reciprocal translocation. Pachytene analysis of F₁ hybrids helped construct chromosome maps based on chromosome length and euchromatin and heterochromatin distribution. Chromosomes were numbered in descending order of 1–20. Pachytene chromosomes in soybean showed heterochromatin distribution on either side of the centromeres. Pachytene analysis revealed small structural differences for chromosomes 6 and 11 which were not detected at diakinesis and metaphase I. This study suggests that G. max and G. soja carry similar genomes and validates the previously assigned genome symbol GG.Research supported in part by the Illinois Agricultural Experiment Station and U.S. Department of Agriculture Competitive Research Grant (85-CRCR-1-1616)     

野生大豆花发育相关基因GsLFY的功能研究

LEAFY/FLORICAULA (LFY/FLO)是植物特有的转录因子家族,在控制花器官的诱导与发育中起着重要的作用,但是与野生大豆花发育相关的LFY/FLO同源基因的研究尚未见报道。本研究从野生大豆中克隆获得1个LFY同源基因,命名为GsLFY,该基因CDS全长1224 bp,包含完整的开放阅读框,编码407个氨基酸。利用实时荧光定量PCR技术对GsLFY在不同组织中的表达情况进行了分析,结果显示GsLFY在根、花以及种子中表达,在茎、叶、茎尖中不表达; 在花发育的四轮不同器官中(萼片、花瓣、心皮和雄蕊)进行实时荧光定量PCR,结果显示GsLFY在花萼和雄蕊中表达,在花瓣和心皮中不表达。酵母单杂交实验结果显示,GsLFY具有转录激活活性。拟南芥原生质体瞬时表达结果表明,GsLFY定位于细胞核中。转GsLFY基因烟草植株开花期比对照提前约29 天,这为通过分子育种的方法获得花期改变的大豆新品种提供了基因资源和理论基础。     

干旱胁迫下外源水杨酸对野生大豆生理特性的影响

目的:研究不同浓度的水杨酸对10％聚乙二醇(PEG-6000)胁迫下野生大豆生理特性的影响.方法:以野生大豆为试验材料,采用盆栽土培法,研究野生大豆幼苗抗氧化酶活性,渗透调解物质及膜质过氧化变化规律.结果:1.5 mmol/L水杨酸明显提高了PEG胁迫下野生大豆过氧化物酶(POD)、超氧化物歧化酶(SOD),减轻了膜脂...     

野生大豆盐碱胁迫相关GsTIFY11b的克隆与功能分析

以耐盐碱野生大豆(Glycine soja L.G07256)为材料,采用同源克隆方法和RT-PCR技术获得一个TIFY类基因的全长cDNA(命名为GsTIFY11b)。进化树分析表明,与其他物种相比,GsTIFY11b与拟南芥的AtTIFY11a基因相似性最高,达到56%;序列分析表明GsTIFY11b蛋白除具有TIFY保守结构域外,还具有一个N端保守结构域和一个C端保守的Jas结构域;实时荧光定量PCR结果显示该基因受盐和碱胁迫诱导表达;将GsTIFY11b转化拟南芥来验证其耐盐碱功能,获得两个转基因纯合体株系,盐碱胁迫分析结果表明,GsTIFY11b的超量表达没能提高拟南芥对盐碱胁迫的耐性,并且与野生型相比,转基因植株在种子萌发期和苗期表现出对盐胁迫更加敏感。盐胁迫信号通路相关marker基因在转基因拟南芥中的表达特性分析表明,GsTIFY11b可以调控RD29B、KIN1、DREB等基因的转录。在洋葱表皮细胞中瞬时表达GsTIFY11b-GFP融合蛋白的结果表明,GsTIFY11b定位于细胞核中。上述结果表明,该基因在细胞核中起着转录调节子的作用,可能是通过调控盐胁迫信号通路中关键基因的表达来改变植物对盐胁迫的耐受性。     

野生大豆盐碱胁迫相关GsTIFY11b的克隆与功能分析

以耐盐碱野生大豆(Glycine soja L.G07256)为材料, 采用同源克隆方法和RT-PCR技术获得一个TIFY 类基因的全长cDNA(命名为GsTIFY11b)。进化树分析表明, 与其他物种相比, GsTIFY11b与拟南芥的AtTIFY11a基因相似性最高, 达到56%; 序列分析表明GsTIFY11b蛋白除具有 TIFY保守结构域外, 还具有一个N端保守结构域和一个C端保守的Jas结构域; 实时荧光定量PCR结果显示该基因受盐和碱胁迫诱导表达; 将GsTIFY11b转化拟南芥来验证其耐盐碱功能, 获得两个转基因纯合体株系, 盐碱胁迫分析结果表明, GsTIFY11b的超量表达没能提高拟南芥对盐碱胁迫的耐性, 并且与野生型相比, 转基因植株在种子萌发期和苗期表现出对盐胁迫更加敏感。盐胁迫信号通路相关marker基因在转基因拟南芥中的表达特性分析表明, GsTIFY11b可以调控RD29B、KIN1、DREB等基因的转录。在洋葱表皮细胞中瞬时表达GsTIFY11b-GFP融合蛋白的结果表明, GsTIFY11b定位于细胞核中。上述结果表明, 该基因在细胞核中起着转录调节子的作用, 可能是通过调控盐胁迫信号通路中关键基因的表达来改变植物对盐胁迫的耐受性。     

一年生野生大豆根系及胚轴皮层通气组织发育观察

用石蜡切片法对一年生野生大豆(Glycine sojaSieb.et Zucc.)的通气组织发育过程进行观测研究,以探讨其耐湿机制.研究表明:(1)在正常生长条件下,一年生野生大豆播种3~5周后主根开始形成皮层通气组织,经过2周左右基本形成根系与茎贯通的通气道;皮层通气组织首先在根茎连接点上下开始形成,再向根尖方向和茎扩展.(2)来自不同生态环境的一年生野生大豆开始形成皮层通气组织的部位相同,但开始产生通气组织的时间存在差异.     

Genetic diversity of nodulating and non-nodulating rhizobia associated with wild soybean (Glycine soja Sieb. & Zucc.) in different ecoregions of China

Microorganisms are important in soil development, inputs and biogeochemical cycling of nutrients and organic matter during early stages of ecosystem development, but little is known about their diversity, distribution, and function in relation to the chemical and physical changes associated with the progress of succession. In this study, we characterized the community structure and activity of nitrogen-fixing microbes during primary succession on a copper tailings. Terminal fragment length polymorphism (T-RFLP) and clone sequencing of nifH genes indicated that different N(2) -fixing communities developed under primary succession. Phylogenetic analysis revealed a diversity of nifH sequences that were mostly novel, and many of these could be assigned to the taxonomic divisions Proteobacteria, Cyanobacteria, and Firmicutes. Members of the Cyanobacteria, mostly affiliated with Nostocales or not closely related to any known organisms, were detected exclusively in the biological soil crusts and represented a substantial fraction of the respective diazotrophic communities. Quantitative PCR (and statistical analyses) revealed that, overall, copy number of nifH sequences increased with progressing succession and correlated with changes in physiochemical properties (including elementary elements such as carbon and nitrogen) and the recorded nitrogenase activities of the tailings. Our study provides an initial insight into the biodiversity and community structure evolution of N(2) -fixing microorganisms in ecological succession of mine tailings.