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。     

转基因甘蔗BtG-2的T-DNA侧翼序列分析及其转化事件特异性检测

转基因甘蔗BtG-2是利用农杆菌介导法把Cry1Ac-2A-gna融合抗虫基因导入‘新台糖22号’的转基因甘蔗株系,具有良好的抗虫特性和农艺性状。为了明确转基因甘蔗BtG-2的分子特征及其检测方法,推进其生物安全性评价工作,以BtG-2的T2代为研究材料,利用Southern杂交检测外源基因在转基因甘蔗基因组内的拷贝数;利用染色体步移技术分离外源基因在甘蔗基因组中插入位点的侧翼序列,并建立了该转化体高效灵敏的特异性PCR检测方法。结果表明：Southern杂交检测证明外源T-DNA以单拷贝方式插入BtG-2株系;经过3次的热不对称巢式PCR扩增,获得外源基因T-DNA左边侧翼序列984 bp和右边侧翼序列705 bp;以这2个序列和相应的T-DNA的左右端序列分别设计3对检测引物对,建立了BtG-2株系的转化事件特异性PCR检测方法,扩增效率最高的引物对LS011/LA451和RS160/RA588分别扩增到440 bp和428 bp的特异片段。其中T-DNA左侧设计的LS011/LA451检测引物对扩增的灵敏度高、特异性强,能够在甘蔗BtG-2基因组DNA相对含量为0.1%的模板中检测出转基因目的成分,相当于9个单倍体基因组拷贝数。本研究完成了转基因株系BtG-2的分子特征及其转化事件特异性检测,为该转基因甘蔗及其衍生产品的检测和身份识别提供技术依据。     

甘蔗品种资源图像信息服务系统简介

采用 Foxpro5 .0 for windows开发出甘蔗品种资源图像信息服务系统 ,本系统由信息数据管理和咨询服务两大模块构成 ,本文主要介绍了甘蔗品种资源图像信息服务系统的运行环境、主要功能及特点     

制糖废水处理方法研究进展

制糖废水是一种高浓度的有机废水 ,如何高效处理是环境工程研究的一个重点。本文就其国外最新的研究成果进行了总结     

Detection, identification and significance of phytoplasmas in grasses in northern Australia

Sugarcane yields have been severely reduced by white leaf and grassy shoot phytoplasma diseases in many parts of Asia. Australian sugarcane crops are not known to be affected by these diseases, but plant pathogenic phytoplasmas found in other introduced and native grasses in northern Australia could pose a serious threat to the Australian sugarcane industry. To further evaluate this threat, leaves from plants of 20 grass species, with and without symptoms, were collected during field surveys in northern Australia and tested to determine whether phytoplasmas were present and whether symptoms were reliable indicators of phytoplasma presence. Molecular tools were used to detect and characterize phytoplasmas. Four different phytoplasmas were found in seven grass species known to grow near healthy sugarcane crops. All the phytoplasmas were closely related to sugarcane white leaf phytoplasma (SCWL), one of the phytoplasmas that causes disease in sugarcane in Asia. Four of the host plant species and two of the phytoplasmas were new records. The relationship between symptoms and phytoplasma presence was poor. Because some plants with symptoms tested negative for phytoplasmas, a series of surveys was carried out in which flowers, leaves, roots and stems of two known host plant species, Whiteochloa cymbiformis and Sorghum stipoideum, were tested separately on nine occasions during two wet seasons. This was done to investigate the distribution of phytoplasmas within plants over time. Results showed that spatial and temporal variation of phytoplasmas occurred in these two host plant species. Hence, evaluation of disease distribution within a region requires repeated testing of all plant parts from plants without symptoms, as well as those with symptoms. To date, there is no report of a vector capable of transmitting to Australian sugarcane the phytoplasmas found in grasses in this study. If one is present, or occurs in the future, then native and introduced grasses could constitute a large reservoir of phytoplasma for vectors to draw on. This work provides an early warning for the sugarcane industry that the potential for infection exists.     

全国甘蔗野生种质资源的采集和考察

１９８５～１９９３年在全国进行了甘蔗野生种质资源的采集和考察。采集到甘蔗亚族（Ｓａｃｃｈａｒｉｎａｅ）内９个属１８个种的无性系共８２４份，种植在云南农业大学校园内（昆明黑龙潭），建立了甘蔗资源圃。考察了甘蔗属（Ｓａｃｃｈａｒｕｍ）、蔗茅属（Ｅｒｉａｎｔｈｕｓ）、河八王属（Ｎａｒｅｎｇａ）和芒属（Ｍｉｓｃａｎｔｈｕｓ）中主要野生种的地理分布、开花期、形态特征、繁殖方式、生态适应性、锤度和种质流失的情况。探明了割手密（Ｓ．ｓｐｏｎｔａｎｅｕｍＬ．）和斑茅（Ｓ．ａｒｕｎｄｉｎａｃｅｕｍＲｅｔｚ）的开花期与原产地纬度和海拔的相关，探索了割手密与蔗茅（Ｅ．ｆｕｌｖｕｓＮｅｅｓ）茎秆锤度与地理分布的联系。     

A streak disease of pearl millet caused by a leafhopper-transmitted geminivirus

The cause of a streak disease of pearl millet (Pennisetum glaucum), originating from Nigeria, has been attributed to a geminivirus belonging to the African streak virus cluster. A full-length, infectious clone of the virus was obtained which was transmissible by the vectorCicadulina mbila (Naudé). Analysis of the complete nucleotide sequence of the coat protein gene of this virus shows it to be most closely related to sugarcane streak virus. The possible evolutionary implications of this finding are discussed.     

快速灵敏的甘蔗线条花叶病毒免疫学检测体系的建立

 甘蔗花叶病广泛存在于我国甘蔗种植区,严重影响甘蔗产业的高质量发展。近年来甘蔗线条花叶病毒在蔗区肆虐,尽管针对其的血清学检测技术已经建立,但是快速、准确、高通量的检测方法亟待发掘。本研究制备了^SCSMVCP的抗血清,特异性高,与引起甘蔗花叶病的另两种病原 (高粱花叶病毒和甘蔗花叶病毒) 间没有血清学交叉反应。基于该多克隆抗体,建立了直接抗原包被的ELISA、斑点杂交、Western blot和基于多抗的免疫试纸条检测技术。开发的免疫试纸条检测技术能快速、准确、高通量应用于田间病毒鉴定。本文提供了基于血清学的快速、准确、高通量,且便捷的甘蔗线条花叶病毒检测技术,有助于我国蔗区甘蔗花叶病的监测与防控。     

2CZD-1型段茎式甘蔗种植机设计与试验

研制了2CZD-1型段茎式甘蔗种植机,可一次性完成甘蔗种植的开沟、施肥、自动取种、自动排种、覆土和覆膜等工序,适用于宽窄行甘蔗种植。设计了机架、取送种机构、转盘施肥机构、旋耕覆土机构、覆膜机构和液压传动系统等结构,其中取送种机构采用排冗结构实现均匀排种。利用有限元分析软件ANSYS对甘蔗种植机的机架进行模态分析,并通过田间试验获得甘蔗种植机的覆土厚度合格率、种植密度、伤芽率、漏植率及总排肥量稳定性变异系数等性能参数。模拟结果表明,当激励频率为8~80 Hz时,机架容易发生共振,且最大位移可达28.12 mm。田间试验表明,该种植机的种植密度为142325芽/hm^2,覆土厚度合格率为93.6%,伤芽率为2.8%,漏植率为4.7%,种植深度合格率为89.3%,总排肥量稳定性变异系数为5.8%,工作生产率为0.32 hm2/h,符合甘蔗种植机的设计要求,能够显著提高甘蔗种植的效率。     

不同生态环境下甘蔗组合的生长表现和适应性

以2017—2018年组配的119个甘蔗组合为材料,分别在广西的南宁、崇左、来宾同时开展组合评价试验,基于锤重性状进行联合方差分析和遗传参数估计,并采用回归分析和AMMI模型对组合进行稳定性分析。结果表明:不同的组合、环境及组合与环境互作在锤重间的差异均达到了极显著水平(P0.01);3个试验点锤重的广义遗传率均属于中等偏低水平;崇左、来宾试验点的锤重变异系数较大,南宁试验点的锤重变异系数较小;643、404、575、972、636、144、YC95、1470、755、409、701、832、YC37、579等组合表现出高产、稳产和适应性广的特性;结合锤重和组合入选率综合分析,组合449、YC127、796、YC44、533、570、YC123、391、546、403、YC90、252在南宁试验点的表现为锤重和组合入选率较高;组合643、212、YC61、432、903、YC95、YC44、368、YC83、YC127、YC112、701、411、YC90、YC123在崇左试验点的表现为锤重和组合入选率较高;组合643、404、449、144、403、YC48在来宾试验点的表现为锤重和组合入选率较高。