唐菖蒲GhAOS基因的克隆与表达

丙二烯氧合酶(AOS)是茉莉酸(JA)生物合成途径中的关键酶,为深入研究AOS基因在唐菖蒲茉莉酸生物合成途径中的作用及唐菖蒲球茎膨大的分子机制,以唐菖蒲品种‘Rose Supreme'的球茎为试材,采用RT-PCR和RACE技术,克隆到了一个GhAOS的cDNA序列,序列内部含有一个1 533 bp的开放阅读框(ORF...     

甘蓝型油菜BnaLPAT2基因在拟南芥中的种子特异性表达及过表达分析

以植物双元表达载体pBI121为骨架,将其改造成BnaLPAT2的过表达载体(p35S-BnaLPAT2)和种子特异性表达载体(pNapin-BnaLPAT2).采用冻融法将重组质粒分别转入农杆菌感受态细胞(GV3101)中,通过农杆菌介导法转化拟南芥,分别获得阳性植株.对转基因植株种子的脂肪酸成分进行分析,结果表明:...     

唐菖蒲的生物学特性及高产栽培技术

介绍唐菖蒲的生物学特性．并从选地整地、田间管理、病虫害防治、收获加工等方面总结了其高产栽培技术，以供参考。     

柑橘和拟南芥 RIN4基因的过表达载体构建及其对沙田柚的 DNA 转化

用逆转承 cDNA 聚合酶链式扩增（RT‐PCR）方法,分别从克里曼丁橘和拟南芥中克隆出 RIN4基因,连接到表达载体 pFGC5941,构建了柑橘 CcRIN4和拟南芥 A tRIN4基因的过表达载体,并成功转入 EHA105农杆菌。通过农杆菌介导法将过表达载体转入沙田柚,获得柑橘和拟南芥 RIN4基因的转基因沙田柚植株各6株。     

柑橘和拟南芥RIN4基因的过表达载体构建及其对沙田柚的DNA转化

用逆转承-cDNA聚合酶链式扩增(RT-PCR)方法 ,分别从克里曼丁橘和拟南芥中克隆出RIN4基因,连接到表达载体pFGC5941,构建了柑橘CcRIN4和拟南芥AtRIN4基因的过表达载体,并成功转入EHA105农杆菌.通过农杆菌介导法将过表达载体转入沙田柚,获得柑橘和拟南芥RIN4基因的转基因沙田柚植株各6株.     

唐菖蒲丙二烯氧化物环化酶基因GhAOC的克隆与表达分析

克隆唐菖蒲AOC基因的全长cDNA序列,并分析该基因的表达模式及其对茉莉酸(Jasmonic acid,JA)生物合成的影响。以唐菖蒲栽培品种‘Rose Supreme’的球茎为试材,利用RT-PCR和RACE技术克隆AOC基因的全长cDNA序列;利用基因枪方法进行GhAOC基因的亚细胞定位分析;运用实时荧光定量PCR技术分析GhAOC基因的表达模式。克隆了1个全长为898bp的茉莉酸生物合成关键酶丙二烯氧化物环化酶(allene oxide cyclase,AOC)基因的cDNA序列,命名为GhAOC。该序列含有1个735bp的开放阅读框(ORF),编码244个氨基酸,推导的蛋白质分子量为26.52ku。亚细胞定位分析表明GhAOC是叶绿体蛋白。实时荧光定量RT-PCR表达分析显示,该基因在唐菖蒲叶、花、根、匍匐茎、新球茎和籽球上都表达,其中在新球茎和籽球中表达量最高;经过0.1～0.5mmol/L的MJ(methyl jasmonate,茉莉酸甲酯)处理后,逐渐提高了GhAOC基因在球茎中的表达量和内源MJ含量。GhAOC基因的表达促进了唐菖蒲茉莉酸的生物合成。     

不同栽培条件下唐菖蒲生长特性比较

采用周径6~8 cm的超级玫瑰、超级红、日本黄等8个品种唐菖蒲种球作为试验材料,春季、冬季分别种植于露地和日光温室中,进行生长特性比较。结果表明,相同唐菖蒲品种在日光温室内生产出的切花,其茎秆要高于露地栽培,花朵多数比露地栽培的大,开花率、花朵数、茎秆粗度等比露地栽培的差;白玉堂、莎莎、蓝精灵、超级红适应性更强,可以用于冬季温室栽培。     

拟南芥PTR3基因过表达载体构建及转基因植株的获得

[目的]构建PTR3基因的过表达载体,并建立PTR3基因过表达植株,为进一步研究PTR3基因在调控重金属胁迫应答中的功能提供理论依据。[方法]提取野生型拟南芥总mRNA,并以反转录的c DNA为模板扩增出PTR3基因CDS全长,通过限制性核酸内切酶酶切、T4 DNA连接酶连接,将该基因连接到带有35S启动子的p BI121载体上。[结果]将重组载体转化至农杆菌GV3101菌株,通过浸花法将重组载体转化到野生型植株中,通过抗性筛选和遗传鉴定获得纯合的PTR3转基因阳性植株。[结论]PTR3过表达载体构建成功并获得转基因阳性植株,为研究PTR3基因的分子功能奠定基础。     

拟南芥miR395d基因的克隆和过表达载体构建及其在油菜中的转化

MicroRNAs(miRNAs)是一类内源性长度约为21～24个核苷酸,在基因表达、生长发育、细胞周期及环境胁迫等方面起着重要作用的单链非编码小分子RNA。研究表明:植物miRNAs通过转录后以切割的方式对其靶基因的表达起着负调控作用。miR395的靶基因分别为ATP硫酸化酶和硫转运体SULTR2;1,两者在硫同化及转运中起着重要的作用。为了进一步探究miR395的功能,本研究利用PCR法从拟南芥中克隆了miR395d前体基因,并与pCAMBIA1304载体连接,构建了miR395d前体基因的过量表达载体,通过根癌农杆菌介导法将其转化到甘蓝型油菜特选4号中,目前已获得了转基因植株。     

唐菖蒲干腐病病原生物学特性

唐菖蒲干腐病也称唐菖蒲枯萎病是唐菖蒲主要病害之一,病原为尖孢镰刀菌唐菖蒲专化型（Fusarium oxysporum fspgtadioli）。生物学特性研究表明,唐菖蒲干腐病菌在淀粉琼脂培养基上菌丝生长最好,该病菌利用的碳源以果糖最好,麦芽糖最差;牛肉膏为菌丝生长最佳氮源,硝酸钾对菌丝生长最差。以酵母浸膏为病菌产孢最佳氮源,氯化铵中产孢最差;在葡萄糖蛋白培养基上产孢最好;最适孢子萌发的碳源为可溶性淀粉,氮源为牛肉膏。pH6～8适宜产孢,pH7产孢最好;在4-35℃条件下菌丝均能生长,25～28℃最适合菌丝生长和产孢;黑暗有利于菌丝生长和产孢。孢子萌发的适温20～30℃,最适28℃,相对湿度（RH）在90％-100％均能萌发,在水滴中萌发率最高,RH低于90％孢子不萌发;萌发最适pH4～5,pH5最好,高于pH10孢子不萌发,黑暗条件下对孢子的萌发有一定的促进作用。     

RAPD Analysis of Twelve General Species of Gladiolus hybridus Hort

Gladiolus hybridus Hort., also named Chang or- chid, gladiola, Shi Yang jin and Flat Zhulian, is a perennial bulb plant of Iridaceae Gladiolus and one of the famous four cutting flowers in the world. Gladiolus hybridus Hort. is an exotic species. There is…     

ISSR Analysis of 26 General Species in Gladiolus hybridus Hort

ISSR analysis is applied in this study to research the classification and genetic relationship of 26 cultivars of G.hybridus Hort.Total 19 arbitrary primers screened from 100 primers were used for further PCR and diversity analysis.A total of 110 ISSR sites were detected with a mean of 5.63 fragments amplified for each primer.A total of 103 polymorphic DNA fragments were detected among all the 110 amplified fragments,which accounted for a high level of 93.6%of all and could be used to identify different cultivars.The results revealed that the idioplasm resource of G.hybridus Hort cutting flower cultivar had a narrow genetic basis on molecular level,and certain genetic relationship existed in summer large-flowers of G.hybridus Hort varieties.     

RAPD Analysis of M_1 Generation of Gladiolus hybridus Hort Treated by EMS

Gladiolus hybridus Hort is one of the world＇s famous cutting flowers. It is very popular because of the big size, bright color, various shape, and long period of bloom. New species should be cultivated in order to meet the consumers＇ need of asking for the new. Among the technologies of cultivating new species of flowers, mutagenic breeding is a shortcut. This study treated corm bud of G. hybridus Hort with EMS of different consistency. Then M1 after treated was analyzed by RAPD. The result showed that EMS was a very effective mutagenic agent for the corm bud of G. hybridus Hort. With the increase of consistency, the mutagenic range increased first, then decreased, among which 0.6% EMS treatment had the biggest influence. However, with the same EMS consistency, there was not close relevancy between the amount of mutagenic agent and the divergence of plant＇s genomes, which offered a molecular basis for selecting plants with good mutation.     

ISSR Analysis of M_1 Generation of Gladiolus hybridus Hort Treated by EMS

Gladiolus hybridus Hort is one of famous cutting flowers,being famous of big size,bright color,various shapes and long bloom period.New species should be cultivated in order to meet consumers' needs.Mutagenic breeding is a shortcut to cultivate new species of flowers.In this study,corm bud of G.hybridus Hort was treated with different concentrations of EMS.Then M 1 generation was analyzed by ISSR.Results showed that EMS was a very effective mutagenic agent for the corm bud of G.hybridus Hort.With the increa...     

Research on protoplast preparation and culture in Gladiolus hybridus Hort.

Protoplasts prepared from the aseptic leaves of Rose Supreme in Gladiolus hybridus Hort. were cultured. The yield of protoplasts with vigour was the highest under the following conditions: 1.5% cellulase Onzuka RS, 0.5% pectinase Y-23, 0.6 mol· L~(-1) mannitol, pH 5.8, digestion time 2 h, and temperature 27℃. After isolation and purification, protoplasts were cultured in solid and liquid combined medium, which included KM8P, 0.6 mol· L~(-1) mannitol, 1.0 mg· L~(-1) 2, 4-D, 0.2 mg· L~(-1) NAA and 0.2 mg· L~(-1) KT. The division of the protoplasts first emerged in medium after cultured 4 days, and emerged only a few times.     

剑兰叶斑病菌鉴定及其生物学特性研究

【目的】叶斑病是制约广东省台山市剑兰产业的重要因素,对病原菌进行分类鉴定、生物学特性和杀菌剂敏感性研究,旨在为病害防控提供参考。【方法】基于病菌致病性、形态特征和 rDNA-ITS 序列分析确定病菌分类地位,采用菌丝生长速率法和镜检孢子量测定温度、pH、光照、碳源、氮源和杀菌剂对病菌生长的影响。【结果】分离纯化获得性状一致的 4 株菌株,致病性测定表明人工接种发病症状与田间症状表现一致。病菌分生孢子梗褐色,多单生,具隔膜,顶端屈膝状,大小为 51.0~80.0 m×4.0~7.6 m。分生孢子褐色、弓形、中间宽两端窄,向一侧弯曲,4 个细胞,大小为 23.5~32.0 m×11.5~16.0 m。基于病菌 rDNA-ITS 序列进行系统进化分析,病菌与唐菖蒲弯孢霉（Curvularia gladioli）聚类到一起,形成明显分支。给合病菌形态特征和 rDNAITS 序列分析确定病原菌为唐菖蒲弯孢霉（Curvularia gladioli Boerema & Hamers）。适宜病菌菌丝生长和产孢的温度范围为 26~30 ℃,pH 范围为 5.0~7.0;光照促进菌丝生长,黑暗利于产孢;碳源木糖、葡萄糖和有机氮源牛肉膏适宜菌丝生长和孢子产生。咪鲜胺和代森锰锌对菌丝生长抑制作用最强,EC50 分别为 1.23、2.81 g/mL。【结论】广东省台山市剑兰叶斑病病原菌为菌唐菖蒲弯孢霉,适宜生长温度为 26~30 ℃,pH 为 5.0~7.0,对咪鲜胺和代森锰锌敏感。     

唐菖蒲冬季开花技术

本文围绕唐菖蒲花期调节问题进行研究。研究认为，实现唐菖冬季开化的主要技术环节有：种球选择，种球冷藏；营养钵过渡种植；掌握种植时间；适时保温，补充光照和加强田间管理采取上述综合技术措施能使唐菖蒲在１１月至翌年２月间开花，开花率高达９０％，切花品质良好。     

PP333对唐菖蒲试管苗增殖和生根的影响

以唐菖蒲“超级玫瑰”(Rose supreme)的茎尖为试验材料,将MS 0.1 mg/L NAA 0.4 mg/L16-BA作为增殖培养基,1/2MS 1.5 mg/L IBA 0.01 mg/L NAA作为生根培养基,添加不同浓度PP333,结果表明:增殖培养基中添加1.0 mg/L PP333对唐菖蒲试管苗增殖生长有促进作用,减少超度含水态苗的发生;生根培养基中添加0.1 mg/L PP333对唐菖蒲试管苗生根壮苗有良好效应。     

唐菖蒲球茎中抗真菌物质活性测定

在从唐菖蒲球茎中提取抗真菌活性生物大分子的过程中,利用木霉和样品水溶液在相应培养基上共培养,观察其抑菌效果,指导提取、纯化和筛选,并将分离纯化得到的纯生物大分子多糖与几种致人体皮肤病病菌进行抗菌谱测定.结果显示:唐菖蒲抗真菌多糖具有较宽的抗菌谱和较高的抗菌活性.