大花蕙兰原生质体分离纯化

以大花蕙兰的叶片、根为试材,分析不同酶液组合、摇床转速、酶解温度、材料酶液比、酶解时间等因素对其原生质体分离的影响。大花蕙兰叶片、根经过90 min质壁分离预处理后,在温度28 ℃,于摇床上50 r·min^-1振荡酶解6 h,叶、根分别在酶解液为纤维素酶2.0%＋果胶酶0.1%＋离析酶0.2%＋崩溃酶0.2%和纤维素酶2.0%＋果胶酶0.3%＋离析酶0.1%＋崩溃酶0.1%中得到最高产量的原生质体。     

The effect of ethylene inhibitors （AgNO3, AVG）, an ethyleneliberating compound （CEPA） and aeration on the formation of protocorm-like bodies of hybrid Cymbidium （Orchidaceae）

Protocorm-like bodies （PLBs） or thin cell layers （TCLs） derived from PLBs of hybrid Cymbidium Twilight Moon ＇Day Light＇ can induce new or neo-PLBs on Teixeira Cymbidium （TC） medium, which contains 0.1 mg/L a- naphthaleneacetic acid, 0.1 mg/L kinetin, 2 g/L tryptone and 20 g/L sucrose, and is solidified with 8 g/L Bacto agar. This study aimed to assess the response ofneo-PLB formation to an ethylene-liberating compound （2-chloroethylphosphonic acid （CEPA））, to two ethylene inhibitors （silver nitrate （AgNO3） and aminoethoxyvinylglycine （AVG））, and to aeration （made possible by using Milliseal~ or autoclaved filter paper）. AgNO3 at 1 or 2 mg/L in TC medium significantly increased the fresh weight of PLBs while 1 mg/L of AgNO3 also showed a significant increase in the number ofneo-PLB from both half-PLBs and from tTCLs. In contrast, AVG and CEPA inhibited neo-PLB formation. Neo-PLB formation from half-PLB or TCL explants in the presence of aeration resulted in significantly lower neo-PLB weight. The use of AgNO3 and aeration are alternative means to mass produce neo-PLBs for micropropagation purposes.     

墨兰幼叶和成熟叶不同部位叶绿体超微结构和光合作用

墨兰试管苗植株成熟叶片叶绿体基粒较发达,类囊本膜垛叠较紧密。幼叶叶绿体中少有亲锇颗粒,成熟叶的叶绿体中往往既有亲锇颗粒又有淀粉粒。幼叶中基粒数目比成熟叶的少,叶绿体也比成熟叶的小。幼叶的光合放氧速率比成熟叶的低。幼叶中叶尖部叶绿体最大而叶基部最小,但叶尖部的光合放氧速率比叶基部小。成熟叶中叶绿体大小及光合放氧速率区别不明显。通过对各部位叶绿素含量的测定发现,叶绿素含量与光合放氧速率之间没有正相关性     

Some suggestions on the geographical origin of the central, west and north European synanthropic species of Papaver L

KADEREIT, J. W., 1990. Some suggestions on the geographical origin of the central, west and north European synanthropic species of Papaver L . Papaver rhoeas, P. dubium subsp. dubium and subsp. lecoqii, P. argemone subsp. argemone and P. hybridum are considered to be introduced in central, western and northern Europe. An analysis of the geographical distribution of the closest relatives of these five taxa leads to hypotheses on their geographical origin. It is suggested that P. rhoeas originates from the eastern border of the Mediterranean, where it might have differentiated only under human influence. The origin of P. dubium subsp. lecoqii is suggested to lie in S.E. Europe or west Anatolia, and that of subsp. dubium in S.E. central Europe. Papaver argemone subsp. argemone might have differentiated from two related subspecies in west Anatolia. No suggestions can be made with respect to P. hybridum , which is taxonomically isolated. The assumptions necessary for hypotheses on the geographical origin of synanthropically widespread taxa are briefly discussed.     

大花蕙兰基因组DNA提取及RAPD反应条件探索

用SDS、CTAB和改良CTAB法分别提取大花蕙兰叶片基因组DNA,发现以改良CTAB法提取的DNA纯度高,产率也较高。以5’-GGTGCTCCGT-3(’BA440)为随机引物,并以大花蕙兰品‘种金杯(’Cymbidiumhybridiumcv.HiroshimaGoldenCu“pSunnyMoon”)的DNA为模板,对RAPD(RandomAmplifiedPolymorphicDNA)反应体系进行了优化研究,结果表明,25μl反应体系中,Mg2 、TaqDNA聚合酶、引物、模板DNA和dNTP5种主要成分的适宜浓度或用量分别是:2.0mmol/L、1.0U、0.20μmol/L、25ng和0.20mmol/L。扩增程序优化为:94℃预变性4min;94℃变性0.5min,37℃退火1min,72℃延伸1min,40个循环;最后72℃延伸7min。     

蕙兰MADS基因APETALA1/FRUITFULL-like的克隆和时空表达特性

为研究兰花成花转变及花发育的调控机理,利用反转录RT-PCR和RACE的方法,从蕙兰萼片中克隆出一个APETALA1/FRUITFULL-like(AP1/FUL-like)基因,命名为CfAP11,GenBank登录号为JQ031272.1.该基因编码的氨基酸序列与MADS-box蛋白家族中类APl/FUL亚家族中球花石斛FRUITFULL-like具有较高的同源性(84％),系统进化分析表明该蛋白的氨基酸序列与APl/FUL转录因子亚家族中的蛋白聚为一类.生物信息学分析推测表明,该基因编码的蛋白具有MADS保守域和相对保守的K区,二级结构中α-螺旋所占比例较高(58.97％),三级结构与月季、水稻和水仙非常相似.相对荧光定量PCR分析结果表明:CfAPll在根中表达痕量,生殖期比营养期叶片中表达量低、盛花期比花蕾期花葶中表达量高,由此推测,CfAP11可能与蕙兰的成花诱导、花发育有关;并发现CfAP11在盛花期花葶和子房中表达量远高于其他组织,表明其可能以某种机制参与果实的形成过程.     

大花蕙兰组织培养技术研究

以大花蕙兰Cymbidium hybridum侧芽为外植体,以N6、B5、CC、MS为基本培养基,设计无机大量元素、无机微量元素、有机物三因素四水平正交试验,探究大花蕙兰丛生芽增殖的最佳基本培养基、生长调节剂浓度配比以及诱导生根的最佳生长调节剂浓度。结果显示,大花蕙兰丛生芽增殖的最佳培养基组合为B5无机大量元素+ CC无机微量元素+MS有机物+蔗糖30 g·L-1 +琼脂7 g·L-1 +活性炭0.5 g·L-1(pH 5.8～6.0),诱导丛生芽增殖的适宜生长调节剂浓度配比为6-BA 4.0 mg·L-1 + NAA 0.2 mg·L-1,诱导生根的最佳生长调节剂浓度为NAA 1.0 mg·L-1。     

单抗免疫斑点法和组织印迹法检测侵染蝴蝶兰的建兰花叶病毒

应用抗建兰花叶病毒(CymMV)的单克隆抗体, 建立了快速检测蝴蝶兰病样的免疫斑点法(Dot-ELISA)和组织印迹法(Tissue blot-ELISA)。CymMV单抗稀释8000倍时, Dot-ELISA可检出病毒粗汁液的最大稀释度为1:10240; Tissue blot-ELISA中样品1次平切后1~5次印迹与Dot-ELISA样品1:80稀释结果相当, 6~8次印迹与Dot-ELISA 1:320稀释结果相当, 前8次印迹均可以得到满意的检测效果。Tissue blot-ELISA的灵敏度略低     

墨兰共生真菌一新种的分离、培养、鉴定及其生物活性

在对产于云南西双版纳的热带兰花菌根进行研究时,发现一分离自墨兰的菌株,菌丝纯白色,具典型的锁状连合,对其子实体的鉴定表明该菌株代表着一新分类单位——兰小菇Mycena orchidicola Fan et Guo sp.nov.同时,对该新种在兰科种子萌发中的作用作了初步探讨,结果表明该菌可侵入兰科种子,促进种子萌发。     

不同消毒和培养基条件对兰花菌根真菌分离的影响

研究了不同的消毒时间和培养基类型对兰花菌根真菌分离效果的影响,以期获得兰花菌根真菌的最佳分离方法.采用了次氯酸钠和酒精作为消毒试剂,发现依次浸没于75 %酒精60 s、2 %次氯酸钠60 和75 %酒精30 s的组合消毒条件下分离效果最佳;选择了3种培养基（PDA、CZA和MEA）,结果表明MEA培养基的分离效果最为稳定.通过组织分离法分离兰花菌根真菌,筛选得到最佳的分离培养条件,旨在为兰花菌根真菌的多样性以及共生效应研究提供研究基础.