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。     

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%。     

Soil CO2 emission in sugarcane management systems

Sugarcane management systems affect soil attributes such as the carbon cycle. This fact has stimulated the sugar and alcohol industry to refine the sugarcane production systems by replacing the pre-harvest burning (PB) and manual harvest with mechanized harvesting followed by residue deposition. The aim of this study was to evaluate different management systems with respect to C cycling carbon dioxide and soil parameters (chemical, physical and biological) which were determined over the season. Three sugarcane cultivation systems were evaluated at the following periods: (a) PB, (b) 5-year green harvest and (c) 10-year green harvest. The results indicated that CO₂ emission was 36% greater in the 10-year sugarcane green harvest system than in the PB system. The bulk density and macroporosity were the factors that were most affected by the different sugarcane management systems and that significantly influenced soil CO₂ emissions. The principal component analysis showed that soil CO₂ emission was 18% influenced by base saturation (V%) and 14% by pH, especially in the PB area. Additionally, 19% was affected by carbon and macroporosity in the 5- and 10-year green harvest areas, respectively. From our results, it can be concluded that the most CO₂ emissions are in the areas of sugarcane green, this is due to the higher carbon concentration when compared with the area of burning sugarcane. The parameters that most influenced the CO₂ emissions were bulk density, porosity, macroporosity, pH and V%.     

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。     

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.     

甘蔗细茎野生种核心种质构建

以来自我国10个省(市)、自治区的540份甘蔗细茎野生种无性系为材料,根据采集地信息及其在20对SSR引物上的分子标记数据和15个表型性状资料,开展核心种质构建研究。不同取样量(5%、10%、20%、30%、40%、50%、60%、70%、80%和90%)分析表明,10%的取样比例可获得70%以上的变异保留比例,是较好的核心种质取样规模;对5种采集地分组取样策略(等量法、简单比例法、平方根比例法、对数比例法和多样性比例法)和2种无分组取样策略(最大变异保留法和随机抽样法)比较表明,简单比例法获得的核心种质代表性最好,为最优取样策略。最后,在简单比例法取样筛选出的54份核心样品中,又通过定向选择补充了6份具有优异表型性状的材料,构建了含60份无性系的甘蔗细茎野生种核心种质,分子和表型检验都表明本研究所构建的核心种质具有较好的代表性和遗传多样性。     

甘蔗分蘖期间叶面喷施乙烯利后两种内源激素的变化

选用甘蔗品种新台糖16号和新台糖22号为材料,在分蘖前期以0(清水)、100、400 mg/L 3种不同乙烯利浓度进行叶面喷施,定期采样分析了茎尖和伤流液中的生长素(IAA)与细胞分裂素(CTK的含量),同时统计了分蘖率、后期分蘖率。结果表明,100mg/L乙烯利处理后分蘖期茎尖IAA含量减少,根部CTK浓度增加,根部伤流液中CTK/IAA比值变大;促进了甘蔗的分蘖,提高分蘖率,尤以对新台糖22号的效应更大。     

广西高糖割手密遗传多样性的表型分析和RAPD分析

采用表型性状变异分析和聚类分析,以及RAPD分子标记分析的方法,研究21份广西高糖割手密无性系的遗传多样性.结果表明,21份高糖割手密无性系在萌芽率、分蘖率、株高、茎径、小区茎数、单茎重上的变异较大、类型多,基于该6性状的聚类分析可将21份割手密无性系划分为3个类群;对21份割手密无性系进行RAPD分子标记多态性分析,发现这些高糖无性系间亦存在较丰富的等位基因位点变异,基于RAPD分子标记的聚类分析将21份无性系划分为4个类群.本研究表明,21份广西高糖割手密无性系间在DNA水平上和主要性状的表型水平上均存在较为丰富的遗传多样性.     

甘蔗愈伤组织诱导与植株再生研究

以甘蔗粤糖95-168号心叶为材料,分别在添加2,4-D1.5、2.0、2.5、3.0、3.5、4.0、4.5和5.0 mg/L8个处理的MS培养基上培养。试验结果表明,不同浓度的2,4-D对愈伤组织诱导有明显影响,并对愈伤组织分化诱导培养产生影响,其中以2,4-D 2.5 mg/L最佳。幼苗经1/2 MS+NAA1.0 mg/L+AC0.05%培养基诱导生根后成为完整的植株。