中国枝孢属研究Ⅶ

枝孢属Ｃｌａｄｏｓｐｏｒｉｕｍ Ｌｉｎｋ：Ｆｒｉｅｓ一新种：贯众枝孢Ｃ．ｃｙｒｔｏｍｉｉ Ｚ．Ｙ．Ｚｈａｎｇ,Ｈ．Ｈ．Ｐｅｎｇ ｅｔ Ｈ．Ｚｈａｎｇ ｓｐ．ｎｏｖ．和于个新记录种：苏铁枝孢Ｃ．ｃｙｃａｄｉｓ Ｍａｒｃｏｌｏｎｇｏ,茄生枝孢Ｃ．ｓｏｌａｎｉｃｏｌａ Ｖｉｅｇａｓ。新种提供了拉丁文和中文描述及形态图。模式标本收藏于云南农业大学真菌标本室（ＭＨＹＡＵ）。     

中国尾孢菌属及其近似属的研究VI.(英文)

本文报道９种尾孢菌,其中有２个新种：蟹甲草尾孢 Ｃｅｒｃｏｓｐｏｒａ ｃａｃａｌｉａｅ Ｙ． Ｌ． Ｇｕｏ　＆ Ｙ．Ｊｉａｎｇ和菜蓟尾孢Ｃｅｒｃｏｓｐｏｒａ 　ｃｙｎａｒａｅ Ｙ． Ｌ． Ｇｕｏ　＆ Ｙ． Ｊｉａｎｇ,中国新记录种有迪氏尾孢Ｃｅｒｃｏｓｐｏｒａｄｅｍｅｔｒｉｏｎｉａｎａ　 Ｇ． Ｗｉｎｔｅｒ,田菁生尾孢 Ｃｅｒｃｏｓｐｏｒａ　 ｇｌｏｔｈｉｄｉｉｃｏｌａ 　Ｔｒａｃｙ　＆ 　Ｅａｒｌｅ,甘草尾孢 Ｃｅｒｃｏｓｐｏｒａｇｌｙｃｙｒｒｈｉｚａｅ 　（Ｓａｖｕｌｅｓｃｕ　＆　Ｓａｎｄｕ）Ｃｈｕｐｐ,野桐尾孢Ｃｅｒｃｏｓｐｏｒａ　ｍａｌｌｏｔｉ　Ｅｌｌｉｓ　＆ 　Ｅｖｓｒｈ,木薯尾孢Ｃｅｒｃｏｓｐｏｒａ ｍａｎｉｈｏｂａｅ Ｖｉｅｇａｓ,补骨脂尾孢 Ｃｅｒｃｏｓｐｏｒａ ｐｓｏｒａｌｅａｅ－ｂｉｔｕｍｉｎｏｓａｅ Ｓａｖｕｌ．＆ Ｓａｎｄｕ和香豆尾孢Ｃｅｒｃｏｓｐｏｒａ ｔｒａｖｅｒｓｉａｎａ Ｓａｃｃ。文中为新种提供了拉丁文描述并附图,研究的标本保存在中国科学院微生物研究所菌物标本馆（ＨＭＡＳ）。     

枝顶孢属一新记录种

从采自安徽祁门枯牛降自然保护区的一种虫草［Ｃｏｒｄｙｃｅｐｓ．ｓｐ．］的内菌核上分离出一株产生孢梗束的丝孢菌,鉴定为地生枝顶孢,在中国属首次报道。地生枝顶孢Ａｃｒｅｍｏｎｉｕｍ　ｔｅｒｒｉｃｏｌａ（Ｍｉｌｌｅｒ　ｅｔ　ａｌ）Ｇａｍｓ,Ｃｅｐｈａｌｏｓｐｏｒｉｕｍ－ａｒｔｉｇｅ　Ｓｃｈｉｍｍｅｌｐｉｌｚｅ（Ｈｙｐｈｏｍｙｃｅｔｅｓ）,ＧｕｓｔａｖＦｉｓｈｅｒＶｅｒｌａｇ,Ｓｔｕｔｔｇａｒｔ,ｐ．６７,１９７１．Ｆｕｓｉｄｉｕｍ　ｔｅｒｒｉｃｏｌａ　Ｍｉｌｌｅｒ,Ｇｉｄｄｅｎｓ　＆　Ｆｏｓｔｅｒ,Ｍ…     

峨眉山自然保护区的虫草及其它昆虫病原真菌I.

采自四川省峨眉山自然保护区的虫草及其相关昆虫病原真菌,其中除有常见的蛹虫草Ｃｏｒｄｙｃｅｐｓｍｉｌｉｔａｒｉｓ（Ｖｕｉｌｌ）,Ｆｒ粉被虫草（ＣｏｒｄｙｃｅｐｓｐｒｕｉｎｏｓａＰｅｔｃｈ）蝉拟青霉Ｐａｅｃｉｌｏｍｙｃｅｓｃｉｃａｄａｅ（Ｍｉｑｕｅｌ）Ｓａｍｓｏｎ,球孢白僵菌Ｂｅａｕｖｅｒｉａｂａｓｓｉａｎａ（Ｂａｌｓ．）Ｖｕｉｌｌ等昆虫病原真菌外,将主要描述峨眉虫草新种（Ｃｏｒｄｙｃｅｐｓｅｍｅｉｅ     

蛹虫草（Cordceps militaris）无性型的多型现象

在萨氏琼脂和ＰＤＡ上,蛹虫草Ｃｏｒｄｙｃｅｐｓ ｍｉｌｉｔｅｒｉｓ（Ｖｕｉｌｌ）．Ｆｒ无性型,蛹草拟青霉Ｐａｅｃｉｌｏｍｙｃｅｓ ｍｉｌｉｔａｒｉｓ（Ｋｏｂ）．Ｂｒ＆Ｓｍ．的一些单孢子株可自发产生突变,基于多型现象及其它形态特征可分为三种类型：（１）具野生型菌株特征,产孢结构拟青霉型（Ｐａｅｃｉｌｏｍｙｃｅｓ－ｔｙｐｅ）,稳定,菌落通常不自发产生角变,大多数单孢了株属此类群。（２）属此类群的单孢子     

中国热带地区的半知菌Ⅰ.云南尾孢菌属及其近似属的种(英文)

本文报道云南热带地区尾孢菌属及其近似属真菌４５种,其中有一个新种：番木瓜短胖孢（ＣｅｒｃｏｓｐｏｒｉｄｉｕｍｐａｐａｙａｅＹ．Ｌ．Ｇｕｏ,ｓｐ．ｎｏｖ．）,寄生在番木瓜科（Ｃａｒｉｃａｃｅａｅ）番木瓜（ＣａｒｉｃａｐａｐａｙａＬ．）叶上。短胖孢属真菌在番木瓜科植物上是首次报道。文中为新种提供了拉丁文简介、描述并附图。模式标本保藏在中国科学院微生物研究所真菌标本室（ＨＭＡＳ）。寄生在番木瓜（ＣａｒｉｃａｐａｐａｙａＬ．）上的ＣｅｒｃｏｓｐｏｒａｍａｍａｏｎｉｓＶｉｅｇａｓ＆Ｃｈｕｐｐ与本种近似,二者都具有子座,分生孢子梗紧密簇生,但前者分生孢子梗（５～２５×２～３μｍ）和分生孢子（１０～４５×２～３μｍ）均短而窄。番木瓜尾孢（ＣｅｒｃｏｓｐｏｒａｐａｐａｙａｅＨａｎｓｆ．）与本种的区别在于无子座,分生孢子梗色泽深（中度褐色）,分生孢子梗长（５０～２００×３．５～６μｍ）,分生孢子无色,针形,窄（２０～７５×３～５μｍ）。     

中国链格孢属的分类研究 Ⅳ.生于夹竹桃科和番木瓜科植物上的新种和新变种(英文)

报道生于夹竹桃科（Ａｐｏｃｙｎａｃｅａｅ）植物上的链格孢新种２个、新变种２个,即络石链格孢（Ａｌｔｅｒｎａｒｉａ　ｔｒａｃｈｅｌｏｓｐｅｒｍｉ　Ｔ．Ｙ　Ｚｈａｎｇ,Ｘ,Ｆ,Ｌｉｎ　ｅｔ　Ｗ．Ｑ．Ｃｈｅｎ）、细极链格孢络石生变种［Ａ．ｔｅｎｕｉｓｓｉｍａ（Ｎｅｅｓｅｘ　Ｆｒ．）Ｗｉｌｔｓｈｉｒｅｖａｒ．ｔｒａｃｈｅｌｏｓｐｅｒｍｉｃｏｌａ　Ｔ．Ｙ．Ｚｈａｎｇ,Ｘ．Ｆ．ＬｉｎｅｔＷ．Ｑ．Ｃｈｅｎ］、细极链格孢长春花变种［Ａ．ｔｅｎｕｉｓｓｉｍａ（ＮｅｅｓｅｘＦｒ．）Ｗｉｌｔｓｈｉｒｅｖａｒ．ｃａｔｈａｒａｎｔｈｉＴ,Ｙ．ＺｈａｎｇｅｔＸ．Ｆ．Ｌｉｎ］和长春花生链格孢（Ａ.ｃａｔｈａｒａｎｔｈｉｃｏｌａＴ．Ｙ．Ｚｈａｎｇ）,及生于番木瓜科（Ｃａｒｉｃａｃｅａｅ）植物上的番木瓜链格孢（Ａ．ｃａｒｉｃａｅ　Ｔ．Ｙ．Ｚｈａｎｇ,Ｗ．Ｑ．Ｃｈｅｎ　ｅｔ　Ｘ．Ｆ．Ｌｉｎ）．新种和新变种均有拉丁文特征描述,并附绘图．新分类单位的模式标本分别存放在西北农业大学真菌标本室（ＨＭＵＡＢＯ）和山东农业大学植物病理标本室（ＨＳＡＵＰ）．     

竹节花黄斑驳病毒启动了在棉花维管组织中是一个强启动子

用竹节花黄斑驳病毒（Ｃｏｍｍｅｌｉｎａ Ｙｅｌｌｏｗ Ｍｏｔｔｌｅ Ｖｉｒｕｓ,ＣｏＹＭＶ）启动子－ｇｕｓ嵌合基因和ＣａＭＶ３５Ｓ－ｇｕｓ嵌合基因通过根癌土壤杆菌（Ａｇｒｏｂａｃｔｅｒｉｕｍ ｔｕｍｅｆａｃｉｅｎｓ（Ｓｍｉｔｈ ｅｔ Ｔｏｗｎｓｅｎｄ）Ｃｏｎｎ）ＬＢＡ４４０４介导分别转入棉花（Ｇｏｓｓｙｐｉｕｍ ｈｉｒｓｕｔｕｍ Ｌ．ｃｖ．Ｊｉｎｇｍｉａｎ７,Ｊ７Ｇ）,诱导再生了棉花转基因植株。Ｇ     

中国链格孢属的研究Ⅰ

本文报道链格孢属的２个新种：寄生在苦木科（Ｓｉｍａｒｏｕｂａｃｅａｅ）臭椿［Ａｉｌａｎｔｈｕｓ ａｌｔｉｓｓｉｍａ （Ｍｉｌｌ）．Ｓｗｉｎｇｌｅ］上的臭椿链格孢（Ａｌｔｅｒｎａｒｉａ ａｉｌａｎｔｈｉ ｓｐ．ｎｏｖ．）,寄生在桦木科（Ｂｅｔｕｌａｃｅａｅ）黑桦（Ｂｅｔｕｌａ ｄａｈｕｒｉｃａ Ｐａｌｌ．）上的桦木链格孢（Ａ．ｂｅｔｕｌａｅ ｓｐ．ｎｏｖ）,２个新组合：豆链格孢［Ａ．ａｚｕｋｉａｅ（     

弯孢节丛孢和厚皮单顶孢菌株的RAPD分析

对来自不同地方的７个弯孢节丛孢（Ａｒｔｈｒｏｂｏｔｒｙｓ ｍｕｓｉｆｏｒｍｉｓ Ｄｒｅｃｈｓｌｅｒ）的９个厚皮单顶孢（Ｍｏｎａｃｒｏｓｐｏｒｉｕｍ ｅｕｄｅｒｍａｔｕｍ（Ｄｒｅｃｈｓｌｅｒ）Ｓｕｂｒａｍ）标准菌株用４个随机引物（ＯＶＷ－１８,ＯＰＷ－１６,ＯＰＭ－１,ＯＰＫ－１９）进行了随机扩增多态性ＤＮＡ（ＲＡＰＤ）分析。扩增谱带聚类分析结果不能将弯孢节丛孢和厚皮单顶孢的菌株分开,具有附属丝的菌     

Effects of Fungal Elicitors on Cell Growth and Artemisinin Accumulation in Hairy Root Cultures of Artemisia annua

The artemisinin accumulation in the hairy root cultures of Artemisia annua L. was enhanced via a treatment of three fungal elicitors separately ( Verticillium dahliae Kleb., Rhizopus stolonifer (Ehrenb. ex Fr. ) Vuill and Colletotrichum dematium (Pers.) Grove). Among these three elicitors, V. dahliae had the highest inducing efficiency, but none of them manifests any noticeable effects on the cell growth of the hairy root cultures. The artemisinin content of the hairy root cultures treated with V. dahliae elicitor was 1.12 mg/g DW, which was 45% higher than the control (0.77 mg/g DW). The results showed that elicitation was dependent on the elicitor concentration, the incubation period and the physiological stage at which the hairy root cultures were treated. In addition, the authors found that for V. dahliae, the optimum concentration was 0.4 mg carbohydrate per millilitre medium, the strongest response of A. annua hairy root cultures to the elicitation was at the late exponential growth stage, and the highest artemisinin content of the hairy root cultures was on the 4th day post treatment.     

内生真菌Colletotrichum sp. B501的寡糖提取物对黄花蒿发根中青蒿素生物合成的诱导

在黄花蒿(Artemisia annua L.)发根液体培养中,黄花蒿内生炭疽菌(Colletotrichum sp. B501)细胞壁寡糖提取物可促进发根青蒿素的合成.经寡糖诱导子(20 mg/L)处理4 d后,发根青蒿素含量达1.15 mg/g, 比对照高出64.29%.诱导作用与诱导子浓度、作用时间相关.诱导处理1 d后,X射线能谱分析表明黄花蒿发根细胞中Ca2+积累量显著增高,电镜观察发现液泡内出现高电子致密物,具活性氧清除作用的过氧化物酶表现出高活性(6.5 unit*min-1*g-1 FW).诱导处理第三天,细胞核DNA呈梯度条带降解,部分细胞出现程序化死亡.内生菌细胞壁寡糖提取物引起的生理反应有利于细胞中青蒿素的生物合成.     

内生真菌Colletotrichum sp. B501的寡糖提取物对黄花蒿发根中青蒿素生物合成的诱导(英文)

在黄花蒿 (ArtemisiaannuaL .)发根液体培养中 ,黄花蒿内生炭疽菌 (Colletotrichumsp .B5 0 1)细胞壁寡糖提取物可促进发根青蒿素的合成。经寡糖诱导子 (2 0mg/L)处理 4d后 ,发根青蒿素含量达 1.15mg/g ,比对照高出6 4 .2 9%。诱导作用与诱导子浓度、作用时间相关。诱导处理 1d后 ,X射线能谱分析表明黄花蒿发根细胞中Ca2 积累量显著增高 ,电镜观察发现液泡内出现高电子致密物 ,具活性氧清除作用的过氧化物酶表现出高活性 (6 .5unit·min-1·g-1FW)。诱导处理第三天 ,细胞核DNA呈梯度条带降解 ,部分细胞出现程序化死亡。内生菌细胞壁寡糖提取物引起的生理反应有利于细胞中青蒿素的生物合成。     

促进黄花蒿发根青蒿素合成的内生真菌诱导子的制备

应用酸解法对黄花蒿(ArtemisiaannuaL.)内生胶孢炭疽菌(Colletotrichumgloeosporioides)菌丝体进行提取,在黄花蒿发根培养系统中比较了各制备提取物的青蒿素诱导活性。活性提取物经过SephadexG25层析后,部分纯化的内生菌寡糖提取物(MW<2500)可显著促进发根青蒿素的合成,培养23d的发根经诱导子(0.4mg/mL)处理4d后,青蒿素产量可达13.51mg/L,比同期对照产量提高51.63%,诱导作用与诱导子浓度、作用时间相关。内生菌寡糖诱导子的制备和使用,在青蒿素生物技术生产研究中为首次应用。     

Nitric oxide potentiates oligosaccharide-induced artemisinin production in Artemisia annua hairy roots

The purpose of the present study was to characterize the generation of nitric oxide （NO） in Artemisia annua roots induced by an oligosaccharide elicitor （OE） from Fusarium oxysporum mycelium and the potentiation role of NO in the elicitation of artemisinin accumulation. The OE （0.3 mg total sugar/mL） induced a rapid production of NO in cultures, which exhibited a biphasic time course, reaching the first plateau within 1.5 h and the second within 8 h of OE treatment. Artemisinin content in 20-day-old hairy roots was increased from 0.7mg/g dry wt to 1.3 mg/g dry wt by using the OE treatment for 4d. In the absence of OE, the NO donor sodium nitroprusside （SNP） at 10, 50 ~1 and 100 ~1 enhanced the growth of hairy roots, but had no effect on artemisinin synthesis, The combination of SNP with OE increased artemisinin content from 1.2 mg/g drywt to 2.2 mg/g dry wt, whereas the maximum production of artemisinin in cultures was 28.5 mg/L, a twofold increase over the OE treatment alone. The effects of SNP on the OE-induced artemisinin were suppressed strongly by the NO scavenger 2-（4- carboxyphenyl）-4,4,5,5-tetramethylimidazoline-l-oxyl-3-oxide （cPTIO）. The results suggest that NO can strongly potentiate elicitor-induced artemisinin synthesis in A. annua hairy roots.     

Improvement of artemisinin accumulation in hairy root cultures of Artemisia annua L by fungal elicitor

The effect of fungal elicitor, derived from mycelial extracts of Penicillium chysogenum 3446, on artemisinin production in hairy root cultures of Artemisia annua L was studied. Various concentrations of elicitor were added to the culture medium after 18 days. Time course experiments were carried out using a defined concentration of elicitor after 18 days. Various ages of hairy root cultures were elicited using a defined concentration of elicitor for 3 days. Artemisinin production in 21-day hairy root cultures treated with 0.3 mg total sugar/ml medium elicitor for 3 days reached to 549.1 mg/l.     

Ri质粒转化的青蒿发根培养及青蒿素的生物合成

用发根农杆菌(Agrobacterium rhizogenes)转化药用植物青蒿(Artemisia annua L．)并建立了发根体外培养系统。Southern杂交、NPT Ⅱ酶的检测证明Ri质粒的T—DNA转移并整合到植物的核基因组上。在发根培养系统中,检测了青蒿的重要次生代谢物一青蒿素的含量,检测了不同理化因子对发根生长及青蒿素含量的影响。结果表明：光照(日光灯,12h光周期,20001x)有利于次生产物青蒿素的积累。培养基的pH值为5．4。蔗糖浓度为3％不仅促进发根的生长,而且促进青蒿素的积累。低浓度萘乙酸(NAA)对发根生长具有促进作用,但抑制青蒿素的合成。赤霉素GA,对发根的生长及次生产物的合成都具有促进作用,其最适浓度为4．8mg／L。     

Effect of plant growth regulators and elicitors on rhinacanthin accumulation in hairy root cultures of Rhinacanthus nasutus (L.) Kurz

Several biologically active secondary metabolites like anthraquinones, sterols, triterpenes, flavonoids and naphthoquinones are present in Rhinacanthus nasutus. Naphthoquinones are important group of compounds generally known as rhinacanthin (RC) consists of 15 derivatives named RC A–D and G–Q of which RC-C, RC-D and RC-N have various medicinal properties. The individual role of two auxins i.e. indole-3-butyric acid (IBA) and α naphthalene acetic acid (NAA) and two elicitors i.e. methyl jasmonate (MJ) and salicylic acid (SA) in Murashige and Skoog medium on hairy root growth and RC (RC-C, RC-D and RC-N) accumulation was investigated in the present study. Time course study revealed that IBA and NAA at 2.5 μM showed maximum fresh weight (FW) and dry weight (DW) 4 weeks after culture. However, RC production was maximum after 6 weeks of culture on both media. A concentration-dependent response was observed when various concentrations of MJ (2.0, 5.0, 10 and 15 μM) and SA (10, 50, 100 and 150 μM) were supplemented in the medium. On MJ and SA media the FW and DW decreased as the concentration of elicitors increased. However, this decrease was more severe in MJ treated cultures. All the MJ and SA treated cultures showed significantly higher amount of RC-C, RC-D and RC-N in hairy roots harvested 7 days after elicitation as compared to control. Of the two elicitors, MJ was more efficient in inducing RC accumulation than SA. The highest RC content (6.3 mg/g DW RC-C; 1.1 mg/g DW RC-D and 0.61 mg/g DW RC-N) was observed after treatment with 10 μM MJ which was about 1.7-, 2.5- and 3.5-fold higher RC-C, RC-D and RC-N respectively than the control.     

Production of artemisinin by hairy rot cultures of Artemisia annua L in bioreactor

Hairy root cultures of Artemisia annua L were cultivated in four different culture systems: a flask, a bubble column, a modified bubble column and a modified inner-loop airlift bioreactor. The artemisinin contents of hairy root cultures in the bubble column and the modified inner-loop airlift bioreactor were higher than that in the modified bubble column. The growth rate and hairy root distribution in the modified inner-loop airlift bioreactor were better than those in other bioreactors, and dry weight and artemisinin production reached to 26.8 g/L and 536 mg/L after 20 days.     

Enhanced production of phytoestrogenic isoflavones from hairy root cultures of Psoralea corylifolia L. Using elicitation and precursor feeding

The effect of biotic elicitors (yeast extract, chitosan), signaling molecule (salicylic acid), and polyamines (putrescine and spermidine) was studied with respect to isoflavones accumulation in hairy root cultures of Psoralea corylifolia L. Untreated hairy roots (control) accumulated 1.55% dry wt of daidzein and 0.19% dry wt of genistein. In precursor feeding experiment, phenylalanine at 2 mM concentration led to 1.3 fold higher production of daidzein (1.91% dry wt) and genistein (0.27% dry wt). In biotic elicitors, chitosan (2 mg/L) was found to be the most efficient elicitor to induce daidzein (2.78% dry wt) and genistein (0.279% dry wt) levels in hairy roots. Salicylic acid at 1 mM concentration stimulated the maximum accumulation of daidzein (2.2% dry wt) and genistein (0.228% dry wt) 2 days after elicitation. In case of polyamines, putrescine (50 mM) resulted in highest accumulation of daidzein (3.01% dry wt) and genistein (0.227% dry wt) after 5 days of addition. Present results indicated the effectiveness of elicitation and precursor feeding on isoflavones accumulation in hairy roots of P. corylifolia. This is the first report of elicitation on isoflavones production by hairy roots of P. corylifolia.