Molecular detection of sugarcane striate virus and sugarcane white streak virus and their prevalence in the Miami World Collection of sugarcane and related grasses

Viruses in the genus Mastrevirus (family Geminiviridae), including those infecting sugarcane, have natural geographical ranges almost exclusively restricted to Africa and the Indian Ocean islands off the African coast. Only sugarcane white streak virus (SWSV) in Barbados and sugarcane striate virus (SStrV) in Florida and Guadeloupe are known to infect a few sugarcane varieties in the Western Hemisphere. In this study, PCR assays were developed to detect these two viruses in sugarcane. Five hundred and seventy-one DNA samples from Saccharum species and interspecific hybrids from the Miami World Collection of sugarcane and related grasses were tested for the presence of SStrV and SWSV by PCR. No variety was found infected by SWSV but SStrV was detected in 19 varieties. PCR data were confirmed by sequencing amplified fragments (248 bp). These fragments shared 93%–100% nucleotide identity with SStrV sequences from the GenBank database. SStrV isolates were distributed in six phylogenetic groups, including the four strains of the virus. Most varieties infected by SStrV originated from Asia, thus confirming a previous hypothesis stating that this virus originated from this continent. Absence of SStrV in commercial sugarcane in Florida also suggested that this virus has not been spread in this location, while infected plants have been present for several decades.     

基于形态特征和SSR标记的斑茅繁殖策略的研究

本研究以收集自我国7个省的45份野生斑茅(Saccharum arundinaceum)为研究对象,在开放授粉条件下,通过测定斑茅的花粉与胚珠比(Pollen-ovule ratio,P/O)、杂交指数(Outcrossing index,OCI)以及基于简单重复序列(Simple sequence repeats,SSR)分子标记估计交配系统参数这3种方法,探究斑茅繁殖特性及其有性繁殖力的情况,为斑茅资源开发利用、杂交育种及繁殖技术提供基础依据。结果表明:斑茅P/O为5 897,杂交指数OCI为2;采用11对SSR引物对随机取样的15个斑茅半同胞家系共计1 158个子代进行交配系统参数估计,结果显示斑茅种群具有较高的异交率水平(t_m=0.864),多位点异交率和单位点异交率的差值不明显(t_m—t_s=0.012),亲本近交系数F大于0(F=0.318),表明斑茅以异交为主,并存在部分近交。综上所述,本研究初步认为斑茅的繁殖特性为异交为主、自交为辅的混合交配系统模式。     

Energy Productivity Potential Assessment of Saccharum spontaneum L.Germplasm Resources

[Objective]The energy productivity potential of Saccharum spontaneum L.germplasm resources as energy plants was discussed in this paper.[Method]The energy productivity potential of Saccharum spontaneum L.germplasm resources collected from the wild was assessed from the aspects of cellulose,semicellulose,lignin and crude ash contents and caloric value.[Result]The total content of cellulose and semicellulose ranged from 41.75%to 69.13%,the lignin content ranged from 2.16%to 11.75%,the crude ash content ranged from 4.79%to 9.34%and the caloric value ranged from 16.00 to 17.69 MJ/kg among the 30 Saccharum spontaneum L.germplasms.Compared to other plants,Saccharum spontaneum L.had higher contents of cellulose and semicellulose,a higher caloric value but lower contents of lignin and crude ash.[Conclusion]From the perspective of producing fuel ethanol with lignocellulose,Saccharum spontaneum L.has a great potential for development as an energy plant.     

甘蔗热带种金属硫蛋白家族基因的克隆及响应重金属胁迫的表达分析

金属硫蛋白是一类富含巯基的低分子量蛋白,在植物的重金属解毒及细胞氧化还原调控等方面起重要的作用。本研究以甘蔗热带种Badila组培苗为材料,分别测定了其在CdCl2、ZnSO4和CuCl2水溶液培养条件下地上部和地下部的重金属含量,结果显示其对上述3种重金属有较强的耐受与富集能力。继而克隆了ScMT1(登录号为KJ504373)、ScMT2-1-5(登录号为MH191346)和ScMT3(登录号为KJ5043704)3个金属硫蛋白家族基因,它们分别属于植物MT亚家族中的MT1、MT2和MT3型基因。ScMT1含有1个内含子和2个外显子,开放阅读框(Open Reading Frame,ORF)长228 bp,编码75个氨基酸;ScMT2-1-5含有2个内含子和3个外显子,ORF长246 bp,编码81个氨基酸;ScMT3含有1个内含子和2个外显子,ORF长198 bp,编码65个氨基酸。RT-qPCR显示,Cd2+胁迫下,在甘蔗地上部和地下部,ScMT2-1-5均连续显著上调表达,而ScMT1的上调应答出现延迟。ScMT3在地上部的上调应答出现延迟,在地下部呈“扬-抑”趋势,提示甘蔗响应Cd^2+胁迫过程中ScMT2-1-5起更积极的作用,ScMT1参与胁迫后期的分子响应,而ScMT3不起主导作用。Cu^2+胁迫下,地上部ScMT1连续显著上调表达,ScMT2-1-5和ScMT3呈总体上调的表达趋势;地下部,ScMT1和ScMT2-1-5的上调表答均出现延迟,仅在胁迫后期显著上调表达,而ScMT3仅在胁迫前期显著上调表达。该结果提示了ScMT1、ScMT2-1-5和ScMT3在Cu2+胁迫响应过程中的协作关系,三者共同参与了地上部的胁迫响应,其中ScMT1起更积极的作用;此外三者还先后参与了地下部对Cu^2+胁迫的分子响应。Zn^2+胁迫下,ScMT1和ScMT3分别仅在地上部和地下部显著上调表达;ScMT2-1-5在地上部和地下部均呈“扬-抑”的应答趋势;提示了在甘蔗响应Cd^2+胁迫应答过程中ScMT1和ScMT3分别在地上部和地下部起主要作用,ScMT2-1-5参与了胁迫前期的分子响应。ScMT1、ScMT2-1-5和ScMT3在甘蔗不同组织中及在重金属(Cd^2+、Zn^2+或Cu^2+)不同累积水平下呈现出相似或互补的应答特性,提示上述甘蔗MT家族不同成员在重金属解毒及细胞氧化还原调控等方面产生了功能分化,且三者在应对过量Cd^2+、Zn^2+或Cu^2+对甘蔗组织造成伤害的过程中存在时空上的协同作用。该研究为深入理解多倍体植物甘蔗中MT家族各成员基因在重金属耐受过程中的协同作用机制奠定了基础。     

基于主要表型性状的不同采集地斑茅种质资源遗传多样性分析

为提高斑茅种质资源利用效率,本研究对来自不同地区的76份斑茅种质资源的株高、茎径、节间长、叶长、叶宽、锤度6个表型性状进行遗传多样性分析。结果表明,来自不同地区的斑茅,6个表型性状平均变异系数在14.6%~53.4%之间,其中株高和节间长的变异系数相对较高,分别为53.4%和37.7%;锤度和叶长的变异系数相对较低,分别为19.1%和14.6%。株高、茎径、叶宽、叶长、节间长之间表现为极显著正相关,其中株高与节间长的相关系数最大,为0.818;锤度与株高、节间长呈负相关,相关系数分别为-0.232和-0.263。主成分分析表明,前2个主成分方差累计贡献率高达72.58%,其中第1主成分方差贡献率达到51.21%,第2主成分贡献率为21.37%。采用聚类分析方法,不同的种质资源被分为8大类群,第Ⅴ和Ⅵ类群的种质综合性状表现最好,这2个类群是优良的斑茅种质资源,可在甘蔗育种中加以挖掘利用。     

斑茅纤维降解生产燃料酒精的发酵工艺研究

以能源植物斑茅纤维为原料,探讨纤维素糖化液生产燃料酒精的发酵工艺研究,分析了时间、温度、pH、氮源种类、酵母膏用量与酵母用量等因素对斑茅纤维素酒精发酵的影响。结果表明:反应时间72~96h,温度35℃,pH4.8~5.2,酵母膏氮源用量0.8%~1.0%,酵母用量10%~15%时斑茅纤维糖化液的酒精发酵效果最好。通过正交试验确定了最佳发酵条件:时间84h、温度35℃、酵母膏用量1.2%、酵母用量20%,发酵验证酒精值达4.6%(V/V),酒精得率32.62%。     

割手密线粒体DNA的简易快速提取方法

高效快速地分离提取高质量的线粒体DNA(mtDNA)是研究植物线粒体基因及其起源进化的重要前提。为获得高质量的mtDNA进行割手密资源的线粒体基因序列扩增及测序分析,本研究以割手密黄化苗为材料,通过简单差速离心分离得到线粒体,经DNaseⅠ消化处理,去除核DNA杂质得到较纯的线粒体,然后经过5%SDS和蛋白酶K充分裂解线粒体,利用饱和的苯酚/氯仿(1:1)和氯仿两次抽提除去蛋白质,并经过RNase A的消化处理除去RNA,无水乙醇沉淀等一系列操作得到mtDNA。所提取的mtDNA样品经紫外吸收光度测定,琼脂糖凝胶电泳和PCR扩增分析其浓度和纯度,结果表明利用该方法提取的mtDNA纯度高,完全能满足后续PCR分析及分子克隆测序的要求。该方法不仅DNA提取纯度高,操作简单、快速经济,可为今后开展甘蔗及其各野生种的研究提供技术支持。     

Evaluation of resistance to Fusarium head blight in indigenous Japanese species of Agropyron (Elymus)

General (GSI) and specific (SSI) selection indices were constructed for three open-mating populations of sugarcane (Saccharum spp.) seedlings and their ratoon stages. Though specific indices were found to be the most effective in improving mean brix yield in their own source populations, but the GSI was better to SSI in other populations as indicated by higher mean values and more number of superior clones at selection and evaluation stages. This indicated the wider applicability of the GSI over SSI. Inspite of better expected genetic advance through GSI in ratoon crop, the actual gain in brix yield attained, through GSI, was better in seedlings. This revised version was published online in July 2006 with corrections to the Cover Date.     

The effect of accidental selfing on the analysis of a diallel cross with sugar cane

Summary The diallel cross was found to be an unsuitable design for quantitative genetic analyses with sugar cane.Accidental selfing caused substantial inbreeding depression for most characters. Consequently, analysis of a diallel cross without regard to the effect of accidental selfing would be subject to bias, if accidental selfing were common. A selfing model proposed by Dudley (1963) was extended to include differential selfing, and was tested in an experiment in which the percentage of selfing in each cross was estimated visually.Variance components had large standard errors, which prevented firm conclusions from being drawn, but selfing appeared to cause substantial bias in the estimates. Estimates of genetic variance components from a p(p-1) diallel set, using the extended selfing model, were compared with estimates obtained from the standard analyses in which reciprocal or maternal effects should account for the effect of selfing. The estimates were comparable, but the standard analyses were simpler and produced estimates with lower standard errors.When the effects of selfing were minimized, general combining ability (g.c.a.) effects appeared to be much more important than specific combining ability (s.c.a.) effects for most characters.