植物病原真菌产生ABA的初步研究

根据植物病理学和微生物生态学原理,建立了脱落酸产生菌的筛选模型,从不同的植和寄主上分离到３６株真菌,其中有十余株菌株能产生天然的脱落酸,这些真菌分别属于葡萄怨属、青霉属、根霉属和镰孢属。     

脱落酸产生菌的遗传育种

从南京蔬菜研究所植物致病株中分离到３株脱落酸（ＡＢＡ）产量较高菌株,经初步鉴定均为灰葡萄孢。分别命名为Ａ２３,Ａ１０１,Ａ１６０。通过紫外诱变及菌株生长状况比较,选出一株菌Ａ１０１作为出发菌,经紫外,硫酸二乙酯多次诱变,在ＡＢＡ合成代谢关键酶抑制剂平板上定向选育出一株ＡＢＡ高产菌ＵＵＤ－１,该菌固体发酵可产生ＡＢＡ３７４μｇ／ｍｌ,是出发菌株产量的１４倍,且其产量连续培养５代稳定。     

脱落酸产生菌的初步筛选

从三份不同来源的土样中分离出４０株真菌。以脱落酸（±）ＡＢＡ为对照,通过移块法从中筛出１５株脱落酸产生菌,并进一步从中复筛出１０个较优的脱落酸产生菌,用丙酮－乙酸乙酯抽提,证实其中８株能产生脱落酸。通过对菌落培养特征和形态特征的观察,初步鉴定它们分属于萄葡孢属、曲霉属和青霉属。     

龙血树真菌群及其对血竭形成的影响

从柬埔寨龙血树（Ｄｒａｃａｅｎａｃｏｃｈｉｎｃｈｉｎｅｎｓｉｓ）茎杆中分离到３０３株真菌,其中镰刀菌属（Ｆｕｓａｒｉｕｍ）菌株占总分离频率的５２％,其次是短梗霉（Ａｕｒｅｏｂａｓｉｄｉｕｍ）和枝孢霉（Ｃｌａｄｏｓｐｏｒｉｕｍ）。通过活体接种对血竭产生的影响试验表明,对血竭形成起重要作用的真菌主要是禾谷镰刀菌龙血树变种（Ｆ．ｇｒａｍｉｎｕｍｖａｒ．ｄｒａｃａｅｎａ）等４株红色镰刀菌,可使血竭形成量提高６６％－１２０％。     

南极淡水分离出的菌物

本文报道从南极乔治王岛所采１１个水样中分离了同的菌物９８株,初步鉴定出１７属２１种,其中６属１３种为南极新记录。后者是：桃色顶孢Ａｃｒｅｍｏｎｉｕｍ ｐｅｒｓｉｃｉｎｕｍ （Ｎｉｃｏｔ） Ｗ．Ｇａｍｓ,密顶孢Ａ．ｓｔｒｉｃｔｕｍ Ｗ．Ｇａｍｓ,构巢曲霉Ａｓｐｅｒｇｉｌｌｕｓ ｎｉｄｕｌａｎｄｓ（Ｅｉｄａｍ）Ｗｉｎｔｅｒ,聚多曲霉Ａｓｐ。ｓｙｄｏｗｉｉ （Ｂａｉｎｉｅｒ ＆ Ｓａｒｔ）Ｔｈｏｍ ＾Ｃ     

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     

链霉菌S01菌株几丁质醇对植物病原真菌的拮抗作用

纯化后的链霉菌Ｓ０１菌株几丁质酶用环柱法在ＰＤＡ平板上对杨树腐烂病菌、葡萄孢菌（Ｂｏｔｒｙｔｉｓ ｓｐ．ＡＳ３．２６１６）、黄瓜黑腥病菌、辣椒疫病菌、棉花黄萎病菌、立枯丝核菌等植物病原真菌及产黄青霉、啤酒酵母的生长具有明显的抑制作用。在６株植物病原真菌的液体培养物中加入几丁质酶后,菌丝出现扭曲变形、细胞质聚集、外溢等异常现象。在高渗缓冲液中,经几丁质酶处理后,啤酒酵母、葡萄孢菌和产黄青霉的细胞壁受     

丛枝菌根真菌对玉米和棉花内源激素的影响*

在温室盆栽条件下研究了丛枝菌根（Ａｒｂｕｓｃｕｌａｒ　ｍｙｃｏｒｒｈｉｚａｓ,ＡＭ）真菌：ＧｉｇａｓｐｏｒａｒｏｓｅａＮｉｃｏｌ．＆Ｓｃｈｅｎｃｋ、Ｇｌｏｍｕｓ　ｍｏｓｓｅａｅ（Ｎｉｃｏｌ．＆　Ｇｅｒｄ．）Ｇｅｒｄｅｍａｎｎ　＆Ｔｒａｐｐｅ和Ｇｌｏｍｕｓ　ｖｅｒｓｉｆｏｒｍｅ　（Ｋａｒｓｔｅｎ）Ｂｅｒｅｈ对玉米和棉花植株内源激素的影响。结果表明,ＡＭ真菌在正常供水和干旱条件下均能显著提高玉米和棉花植株叶片和根内玉米素、生长素和赤霉素的含量,并降低脱落酸的含量。在植物体内含磷量、生长量及其生长发育阶段等一致、仅存在接种与不接种唯一差异条件下,供试ＡＭ真菌同样能改变玉米和棉花植株内源激素的平衡状况。接种处理植株的脱落酸含量与气孔阻力呈正相关关系。表明玉米和棉花植株抗旱性和生长状况的改善与ＡＭ真菌改变内源激素的平衡状况有关。接种ＡＭ真菌的植株表现较强的抗旱性;其生长量也显著大于不接种的对照。ＧＩ．ｖｅｒｓｉｆｏｒｍｅ的效应最大。     

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）．     

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（     

The biosynthetic pathway to abscisic acid via ionylideneethane in the fungus Botrytis cinerea

The biosynthetic pathway to abscisic acid (ABA) from isopentenyl diphosphate in the fungus, Botrytis cinerea, was investigated. Labeling experiments with (18)O2 and H2(18)O indicated that all oxygen atoms at C-1, -1, -1' and -4' of ABA were derived from molecular oxygen, and not from water. This finding was inconsistent not only with the known carotenoid pathway via oxidative cleavage of carotenoids, but also with the classical direct pathway via cyclization of farnesyl diphosphate. The fungus produced new C15-compounds, 2E,4E-alpha-ionylideneethane and 2Z,4E-alpha-ionylideneethane, along with 2E,4E,6E-allofarnesene and 2Z,4E,6E-allofarnesene, but did not apparently produce carotenoids except for a trace of phytoene. The C15-compounds labeled with 13C were converted to ABA by the fungus, and the incorporation ratio of 2Z,4E-alpha-ionylideneethane was higher than that of 2E,4E-alpha-ionylideneethane. From these results, it was concluded that farnesyl diphosphate was reduced at C-1, desaturated at C-4, and isomerized at C-2 to form 2Z,4E,6E-allofarnesene before being cyclized to 2Z,4E-alpha-ionylideneethane; the ionylideneethane was then oxidized to ABA with molecular oxygen. This direct pathway via ionylideneethane means that the biosynthetic pathway to fungal ABA, not only before but also after isopentenyl diphosphate, differs from that to ABA in plants, since plant ABA is biosynthesized using the non-mevalonate and carotenoid pathways.     

The P450 monooxygenase BcABA1 is essential for abscisic acid biosynthesis in Botrytis cinerea

The phytopathogenic ascomycete Botrytis cinerea is known to produce abscisic acid (ABA), which is thought to be involved in host-pathogen interaction. Biochemical analyses had previously shown that, in contrast to higher plants, the fungal ABA biosynthesis probably does not proceed via carotenoids but involves direct cyclization of farnesyl diphosphate and subsequent oxidation steps. We present here evidence that this "direct" pathway is indeed the only one used by an ABA-overproducing strain of B. cinerea. Targeted inactivation of the gene bccpr1 encoding a cytochrome P450 oxidoreductase reduced the ABA production significantly, proving the involvement of P450 monooxygenases in the pathway. Expression analysis of 28 different putative P450 monooxygenase genes revealed two that were induced under ABA biosynthesis conditions. Targeted inactivation showed that one of these, bcaba1, is essential for ABA biosynthesis: DeltaBcaba1 mutants contained no residual ABA. Thus, bcaba1 represents the first identified fungal ABA biosynthetic gene.     

Biosynthesis of abscisic acid by the direct pathway via ionylideneethane in a fungus, Cercospora cruenta

We examined the biosynthetic pathway of abscisic acid (ABA) after isopentenyl diphosphate in a fungus, Cercospora cruenta. All oxygen atoms at C-1, -1, -1', and -4' of ABA produced by this fungus were labeled with (18)O from (18)O(2). The fungus did not produce the 9Z-carotenoid possessing gamma-ring that is likely a precursor for the carotenoid pathway, but produced new sesquiterpenoids, 2E,4E-gamma-ionylideneethane and 2Z,4E-gamma-ionylideneethane, along with 2E,4E,6E-allofarnesene. The fungus converted these sesquiterpenoids labeled with (13)C to ABA, and the incorporation ratio of 2Z,4E-gamma-ionylideneethane was higher than that of 2E,4E-gamma-ionylideneethane. From these results, we concluded that C. cruenta biosynthesized ABA by the direct pathway via oxidation of ionylideneethane with molecular oxygen following cyclization of allofarnesene. This direct pathway via ionylideneethane in the fungus is consistent with that in Botrytis cinerea, except for the positions of double bonds in the rings of biosynthetic intermediates, suggesting that the pathway is common among ABA-producing fungi.     

Identification of an abscisic acid gene cluster in the grey mold Botrytis cinerea

Like several other phytopathogenic fungi, the ascomycete Botrytis cinerea is known to produce the plant hormone abscisic acid (ABA) in axenic culture. Recently, bcaba1, the first fungal gene involved in ABA biosynthesis, was identified. Neighborhood analysis of bcaba1 revealed three further candidate genes of this pathway: a putative P450 monooxygenase-encoding gene (bcaba2), an open reading frame without significant similarities (bcaba3), and a gene probably coding for a short-chain dehydrogenase/reductase (bcaba4). Targeted inactivation of the genes proved the involvement of BcABA2 and BcABA3 in ABA biosynthesis and suggested a contribution of BcABA4. The close linkage of at least three ABA biosynthetic genes is strong evidence for the presence of an abscisic acid gene cluster in B. cinerea.     

Interaction of abscisic acid and indole-3-acetic acid-producing fungi with Salix leaves

The molds Botrytis cinerea, Cladosporium cladosporioides, and the yeast Aureobasidium pullulans, isolated from the leaves of three short-rotation Salix clones, were found to produce indole-3-acetic acid (IAA). Abscisic acid (ABA) production was detected in B. cinerea. The contents of IAA and ABA in the leaves of the Salix clones and the amounts of fungal propagules in these leaves were also measured, in order to evaluate whether the amounts of plant growth regulators produced by the fungi would make a significant contribution to the hormonal quantities of the leaves. The content of ABA, and to a lesser degree that of IAA, showed a positive correlation with the frequency of infection by the hormone-producing organisms. The amounts of hormone-producing fungi on leaves that bore visible colonies were, however, not sufficiently high to support the claim that either the fungal production of ABA or IAA would significantly contribute to the hormonal contents of the leaves of the Salix clones. It is therefore suggested that the effect of fungal IAA production on plants is limited to the rhizosphere and that B. cinerea, which is a known pathogen, induces ABA production by the mother plant as a response to physiological stress.Abbreviations ABA abscisic acid - ABA-Me abscisic acid methyl ester - GC-MS-SIM gas chromatography-selected ion monitoring-mass spectrometry - IAA indole-3-acetic acid - IAA-Me indole-3-acetic acid methyl ester Author for correspondence.     

The P450 Monooxygenase BcABA1 Is Essential for Abscisic Acid Biosynthesis in Botrytis cinerea

The phytopathogenic ascomycete Botrytis cinerea is known to produce abscisic acid (ABA), which is thought to be involved in host-pathogen interaction. Biochemical analyses had previously shown that, in contrast to higher plants, the fungal ABA biosynthesis probably does not proceed via carotenoids but involves direct cyclization of farnesyl diphosphate and subsequent oxidation steps. We present here evidence that this “direct” pathway is indeed the only one used by an ABA-overproducing strain of B. cinerea. Targeted inactivation of the gene bccpr1 encoding a cytochrome P450 oxidoreductase reduced the ABA production significantly, proving the involvement of P450 monooxygenases in the pathway. Expression analysis of 28 different putative P450 monooxygenase genes revealed two that were induced under ABA biosynthesis conditions. Targeted inactivation showed that one of these, bcaba1, is essential for ABA biosynthesis: ΔBcaba1 mutants contained no residual ABA. Thus, bcaba1 represents the first identified fungal ABA biosynthetic gene.     

Potato cold hardiness development and abscisic acid. I. Conjugated abscisic acid is not the source of the increase in free abscisic acid during potato (Solanum commersonii) cold acclimation

Free and conjugated abscisic acid (ABA) levels in stem-cultured plantlets of potato ( Solanum commersonii Dun, PI 458317) during cold acclimation were measured. The levels of free and conjugated ABA were measured by an enzyme immunoassay (EIA) with rabbit anti-ABA-serum. The use of immunoglobulin G fraction purified from rabbit antiserum and the methylated form of ABA resulted in an improved measuring range (0.01 to 10 pmol ABA) and precision (slope of logit-log plot, −1.35) of EIA, compared to the use of antiserum and free ABA. Estimates of the EIA were consistent with those resulting from a commercial EIA. Under a 4/2°C (day/night) temperature regime, the potato plantlets increased cold hardiness from −5°C (warm-grown control) to −10°C by the 7th day. During the same period, there were two transitory increases in free ABA, the first one three-fold from 1.5 to 5.3 nmol (g dry weight)⁻¹ on the 2nd day and the second one five-fold from 1.5 to 7.6 nmol (g dry weight)⁻¹ on the 6th day. Each increase in ABA concentration was followed by an increase in cold hardiness. There was no significant change in conjugated ABA content (4.2±0.6 nmol [g dry weight]⁻¹) throughout the cold acclimation period. The lack of an interrelationship between levels of free and conjugated ABA suggested that the transitory increase in free ABA during cold acclimation was not a result of the conversion of conjugated ABA. The increase in free ABA due to biosynthesis of ABA during potato cold acclimation is discussed.