Studies on hypersaline-tolerant white-rot fungi IV: effects of Mn 2+ and NH 4 + on manganese peroxidase production and Poly R-478 decolorization by the marine isolate Phlebia sp. MG-60 under saline conditions

Manganese peroxidase (MnP) production by Phlebia sp. MG-60 is strongly regulated by Mn²⁺ and NH₄⁺ at various sea salt incubation conditions. Extra-added Mn²⁺ and NH₄⁺ obviously inhibited MnP production, but sea salts relieved the inhibition partially or completely. Three media were prepared: Kirk medium with addition of 0%–5% sea salts (KS medium), a high level of Mn²⁺ (300mg/l) in KS medium (HM-KS medium), and a high level of NH₄⁺ (430mg/l) in KS medium (HN-KS medium). Without addition of sea salts, the dye Poly R-478 was significantly decolorized by low MnP activity (about 200U/l) and a low level of laccase activity (about 100U/l) in KS and HM-KS media. In the cultures in which laccase activity was almost completely inhibited by 3% and 5% sea salts, MnP activity higher than 400U/l was necessary for Poly R-478 decolorization in all of the three media. We first report the linear correlation of MnP activity and decolorization of Poly R-478 under saline conditions and the effect of laccase on this relation.     

Effect of Lecanicillium muscarium on Eretmocerus sp. nr. furuhashii (Hymenoptera: Aphelinidae), a parasitoid of Bemisia tabaci (Hemiptera: Aleyrodidae)

Pathogenicity of Lecanicillium muscarium, against Eretmocerus sp. nr. furuhashii (Hymenoptera: Aphelinidae), was investigated under laboratory conditions to determine if the fungal infection of the whitefly host can effect the survival, longevity and fecundity of female parasitoid. The results indicated that the number of parasitized larvae surviving a L. muscarium treatment after 6 days of oviposition decreased with increasing concentrations of L. muscarium and in later stages of development (12 days post oviposition) were not affected by fungal application. There were no significant differences on adult parasitoid survivorship after 7 days among all treatments. Maximum survivorship (73.33%) was observed for control and it was minimum (60%) at 1 × 10⁸ conidia/ml. L. muscarium showed a non significant effect on longevity and next offsprings of female parasitoids. The percentage emergence of parasitoids from the whitefly nymphs produced by the females emerged from treated pupae was almost similar. Maximum emergence (69.77%) was observed at 1 × 10⁶ conidia/ml and it was lowest (61.02%) at conidial concentration of 1 × 10⁸ conidia/ml. Maximum longevity of adult Eretmocerus sp. emerging from whitefly nymphs when treated after 12 days of postoviposition was observed for 1 × 10⁵ conidia/ml having a mean value of 5 days whereas the lowest longevity was 4.9 days observed at 1 × 10⁸ conidia/ml. The results mentioned above indicate that the interaction among biocontrol agents is positive to a greater extent with minimum risk hazards.     

桉小卷娥发生规律与防治的研究

桉小卷蛾在我国桉树分布地区普遍发生,危害顶梢和嫩叶,严重影响幼林的高生长。1985～1987年、1991～1993年,先后在海南岛国营上埇林场、广东省雷州林业局对该虫进行了发生规律调查和防治试验,现将结果整理如下。     

林地土壤及其真菌对毛红椿种子发芽及幼苗保存的影响

[目的]通过不同群落毛红椿天然林林地土壤及其真菌对种子萌发和幼苗存活影响的研究,探讨影响毛红椿天然更新的障碍因素。[方法]在江西九连山国家级自然保护区内的3个毛红椿天然优势群落中,分别取距离毛红椿母树3个不同距离(2.5、5.0、7.5 m)处的根区土壤和远离毛红椿母树(25 m以外)的非根区土壤,以非林地土为对照开展室内模拟播种和以根区土壤真菌的水或根系分泌物悬液对毛红椿幼苗进行灌根接种,观察种子萌发、幼苗存活和幼苗感病情况。[结果]各群落土壤中种子发芽均呈先上升后下降规律,且基本在第8 10天达到高峰;林地土与非林地土种子发芽率差异不显著,但林地根区土幼苗的存活率极显著低于非林地土和林地非根区土;距母树3个不同距离根区土的幼苗存活率差异不显著。灌根接种不同处理间幼苗存活率差异极显著,具体表现为接种RS2、RS5根区真菌的幼苗感病率显著高于根系分泌物及无菌水空白对照,且RS2真菌的根系分泌物悬浮液幼苗感病率显著高于其无菌水悬浮液。[结论]毛红椿种子萌发不受土壤环境的影响,但幼苗建成受根区范围内土壤的影响,根系分泌物和致病菌的互作显著降低幼苗保存率。由此推断,毛红椿根区土壤内存在幼苗的潜在致病菌及根系分泌物可增强其致病性。     

Studies on hypersaline-tolerant white-rot fungi III: biobleaching of unbleached kraft pulp by hypersaline-tolerant manganese peroxidase from a marine white rot isolate, <Emphasis Type="Italic">Phlebia </Emphasis>sp. MG-60

 A marine white rot isolate, Phlebia sp. MG-60, secreted lignin peroxidase (LiP), manganese peroxidase (MnP), and laccase under different sea salt incubation conditions. Its MnP production was strongly enhanced by adding 3% sea salts, and the MnP showed high tolerance to sea salts and NaCl. The crude enzyme secreted at 3% sea salt concentration by Phlebia sp. MG-60, in which the main component was MnP (cMnP), was then used to bleach unbleached hardwood kraft pulp (UKP) in vitro. The pulp was brightened 11 points by 4 U of cMnP, and the kappa number was decreased 6 points when only 0.5 mM H₂O₂ was added continuously. When 0.5 mM H₂O₂ (1.22 mg H₂O₂ /g pulp) was added at the initial bleaching, the pulp brightness increased 6 points with a dosage of 4 U of cMnP. When crude MnPs were employed to bleach UKP with organic-free model white-water instead of the Milli Q water usually used, the pulp was brightened 10 and 13 points by 4 and 20 U of cMnP, respectively, and 5 and 6 points by 4 and 20 U of MnP, respectively, of Phanerochaete chrysosporium. Received: September 28, 2001 / Accepted: March 15, 2002 Correspondence to:R. Kondo     

草木樨中华根瘤菌CHW10B溶磷特性及其对南方红豆杉的促生作用

[目的]探讨南方红豆杉根际微生物草木樨中华根瘤菌(Sinorhizobium meliloti)CHW10B的溶磷特性及促生作用。[方法]利用液体发酵试验比较不同时间、碳源、氮源、碳氮比、培养温度等条件下CHW10B菌株的溶磷量,采用温室盆栽法研究该菌株对南方红豆杉生长的影响,并分析该菌株产嗜铁素、精氨酸脱羧酶、ACC脱氨酶及有机酸等成分的能力。[结果]该菌株分别在培养时间4 d、磷酸钙5.00 g·L~(-1)、初始p H值7.0、装液量1/2、NaCl为0.0 g·L~(-1)、温度30℃、碳源为葡萄糖、氮源为硫酸铵、碳氮比100∶1时溶磷能力最强。将该菌株接种于1年生南方红豆杉实生苗360 d后,苗木的地径、苗高、生物量比对照分别增长了19.53%、20.14%、25.39%。该菌株能够产嗜铁素、精氨酸脱羧酶、ACC脱氨酶(0.922 U·mg~(-1))和IAA(8.908 mg·m L~(-1)),并可分泌大量有机酸—葡萄糖酸(5 704.92μg·L~(-1))。[结论]草木樨中华根瘤菌CHW10B溶磷能力强,对南方红豆杉促生作用显著,适用于多种不同酸碱性土壤,可高效应用于南方红豆杉的人工栽培,具有被开发为生物肥料的潜力。     

基于反射光谱指数的锐齿栎叶片色素含量估算

[目的]探讨锐齿栎叶片色素含量和光谱反射率之间的关系,确定无损、快速估算锐齿栎叶片色素含量的敏感反射光谱波段和光谱指数。[方法]在2个样地进行了2年的野外观测试验,于生长季内同步测定了锐齿栎叶片的光谱反射率和不同光合色素含量,分析了350 2 500 nm范围内光谱反射率和敏感光谱指数与叶片色素含量及比率之间的定量关系。[结果]锐齿栎叶片的光谱反射率随叶片色素含量呈明显的规律性变化,与Chl a、Chl b、Chl和Car的含量在可见光的绿光黄光及红边区域表现为显著或极显著的负相关性,与Car/Chl表现为极显著的正相关性;在近红外和短波红外区域,光谱反射率与色素的相关性不及可见光区。本文构建的色素敏感光谱指数ND(705,350)、ND(800,705)、m ND(800,705)和m PRI可以准确地估算锐齿栎叶片的Chl a、Chl b、Chl含量和Car/Chl比率。独立的试验数据检验表明估算值和实测值的拟合关系较好。[结论]ND(705,350)、ND(800,705)、mND(800,705)和mPRI可有效地估算锐齿栎叶片的色素含量及比率。     

Multihydroxylation of ursolic acid by Pestalotiopsis microspora isolated from the medicinal plant Huperzia serrata

The structural modification of ursolic acid by an endophytic fungus Pestalotiopsis microspora, isolated from medicinal plant Huperzia serrata was reported for the first time. The structure diversity was very important for the SAR study of ursolic acid and its derivatives. Incubation of ursolic acid 1 with P. microspora afforded four metabolites: 3-oxo-15α, 30-dihydroxy-urs-12-en-28-oic acid (2), 3β, 15α-dihydroxy-urs-12-en-28-oic acid (3), 3β, 15α, 30- trihydroxy-urs-12-en-28-oic acid (4) and 3,4-seco-ursan-4,30-dihydroxy-12-en-3,28-dioic acid (5). All products were new compounds and their structures elucidation was mainly based on the spectroscopic data.     

花吊丝竹居群遗传多样性的ISSR分析

[目的]研究花吊丝竹居群遗传多样性和遗传结构,为种质资源有效利用和良种选育提供理论指导。[方法]利用12条ISSR引物对48份种质(共3个居群)花吊丝竹居群进行遗传多样性和遗传距离分析。[结果]共检测到124个位点,其中,多态性位点为102个,种质和居群水平上的多态位点百分比(PPB)分别为82. 26%和50. 27%,Ne’基因多样性指数(He)分别为0. 220 4和0. 206 6,Shannon’s信息指数(I)分别为0. 349 4和0. 300 5,表明花吊丝竹居群间存在中等水平的遗传变异。根据Nei’s遗传多样性计算出不同居群间分化水平(Gst)=0. 163 3,表明16. 33%的遗传变异存在于居群间,居群内的遗传变异为83. 67%。居群间的基因流Nm为2. 562 1,表明花吊丝竹居群间存在较大基因流,很大程度减少居群间遗传差异。基于遗传距离的UPGMA聚类结果表明,48份种质可分为3组,3个居群可分为2组,居群间地理距离与亲缘关系无显著相关性。[结论]虽然花吊丝竹主要靠营养生殖来繁衍后代,其居群遗传多样性较丰富,且居群内遗传多样性大于居群间。此外,福建居群遗传多样性明显高于广西和广东地区居群。     

小陇山锐齿栎天然林结构动态分析

[目的]为了解锐齿栎天然林群落动态变化过程。[方法]采用每木定位监测样地重复观测的方法,对甘肃小陇山林区百花林场王安沟营林区内的锐齿栎天然林进行研究,从树种组成及多样性结构、径级结构、空间结构等几方面分析了锐齿栎天然林群落的结构动态特征。[结果]表明:2次调查群落树种组成和优势树种的重要值排序变化不大,有2个稀少种退出群落,死亡林木40株,死亡率8.3%;群落乔木层的物种丰富度和树种空间多样性下降,优势树种的集中性变大,物种个体数目分配的均匀程度下降。林分径级结构由典型的反"J"型分布变化为左偏的单峰状曲线;群落的空间结构没有发生显著变化,林木分布格局仍为随机分布,中林层和上林层林木个体增加,垂直结构更趋复杂;树种隔离程度下降,建群种锐齿栎的优势度增强,膀胱果、白桦和青榨槭种群的优势度下降。[结论]锐齿栎天然林群落组成和结构变化是一个复杂和缓慢的过程,6年间仅发生了一些微小的波动。     

尾叶桉混合菌根营养生理研究

桉树是具有混合菌根（ＥＣＭ和ＶＡＭ）的树种,关于混合菌根营养生理的研究,国内外均未见报道。本文在菌圃接种试验的基础上,研究了尾叶桉苗混合接种ＥＣＭ菌彩色豆马勃和ＶＡＭ菌苏格兰球囊霉对宿主营养生理的影响。结果表明,尾叶桉苗期混合拉种菌根真菌后,对苗木根系营养生理产生了较大的影响。菌根接种有显著提高苗木的根系活力,中混合接种方式分别比未拉种对照苗提高了３１．９０％和２８．６９％,并好一两种单接种处理,     

木麻黄与根系微生物关系研究进展

木麻黄是我国沿海防护林基干林带中的主栽树种,与土壤中的微生物可以形成共生关系,提高自身耐干旱、抗贫瘠、抗盐碱等适应恶劣环境的能力。土壤中的细菌和真菌能够与木麻黄根系形成稳定的相互作用的微生态系统,其中微生物主导的物质代谢循环是植物获取营养的主要来源之一,反之木麻黄的根系分泌物可以为微生物的生长提供养分。文中综述了木麻黄与根系微生物之间相互作用的相关研究,尤其是接种弗兰克氏菌（Frankia）及菌根菌提高木麻黄抗逆性方面的研究进展,对木麻黄与根系微生物体系的促生、抗逆机制进行了探讨和展望。     

基于MaxEnt模型的毛红椿适生区预测

[目的]了解环境因子对毛红椿适生区分布的影响,为其资源保护、引种及其人工林的发展提供参考。[方法]基于MaxEnt模型能利用现存不完整、小样本、离散型分布数据构建物种适生区预测模型,用受试者工作曲线线下面积(AUC)检验预测模型的精度,面积越大精度越高等优点,本研究应用毛红椿在云南的分布数据及1个地型因子和6个气候因子,来构建其适生区分布模型。[结果]毛红椿适生区分布MaxEnt模型平均训练AUC和平均测试AUC分别为0.891、0.885,说明对毛红椿适生区的预测是可靠的;降水量变异系数和最干季度降水量是决定毛红椿适生区分布的主要因子,年均气温变化范围、最冷季度平均气温、最湿季度降水量、最冷季度降水量是次要因子。在当代和未来(2050S、2070S)气候变暖条件下(RCP2.6情景),云南省和全国适生区面积计算结果直观、定量的反应了全球变暖对毛红椿适生区变迁的影响。[结论]预测云南省及全国的毛红椿适生区随全球变暖而小幅萎缩。     

Heptaketides with antiviral activity from three endolichenic fungal strains Nigrospora sp., Alternaria sp. and Phialophora sp

Two new heptaketides, (+)-(2S,3S,4aS)-altenuene (1a) and (-)-(2S,3S,4aR)-isoaltenuene (2a), together with six known compounds, (-)-(2R,3R,4aR)-altenuene (1b), (+)-(2R,3R,4aS)-isoaltenuene (2b), 5'-methoxy-6-methyl-biphenyl-3,4,3'-triol (3), alternariol (4), alternariol-9-methyl ether (5), and 4-hydroxyalternariol-9-methyl ether (6) were isolated from the EtOAc extract of an endolichenic fungal strain Nigrospora sphaerica (No.83-1-1-2). Compounds 1a and 1b were separated from enantiomers 1 by chiral HPLC, and so were 2a and 2b from enantiomers 2. Interestingly, 1-6 were also obtained from other two endolichenic fungal strains Alternaria alternata (No.58-8-4-1) and Phialophora sp. (No.96-1-8-1). The structures of 1-6 were elucidated by means of MS, HR-MS, NMR, and X-ray diffraction. Furthermore, the absolute configurations of 1a-2b were determined by CD experiments and CD calculation. Of these compounds, 4 and 5 showed antiviral activity against herpes simplex virus (HSV) in vitro, with IC(50) values of 13.5 and 21.3 μM, and with selective index (SI) values of 26.5 and 17.1, respectively.     

沙地樟子松天然纯林的结构特征

[目的]为了解红花尔基地区沙地樟子松天然纯林的结构特征,指导沙地樟子松的保护与经营。[方法]在红花尔基地区设置2块100 m×100 m的樟子松天然纯林固定样地,利用样地内每木定位调查数据和分析统计软件进行一元分布及二元分布特征分析。[结果](1)樟子松天然林纯林直径分布为单峰或多峰山状分布,垂直结构简单,只有乔木层和草本层。(2)樟子松天然纯林的林木分布格局为均匀分布,接近随机分布,林木分布格局类型与林分密度无关;林分中樟子松个体竞争激烈,多数单元中林木呈较密集状态。(3)2块样地中随机分布状态下的林木多数为中等密集或比较密集,不同分布状态下的林木优劣性差异较小;低密度樟子松天然纯林中多数密集状态的林木为绝对优势木或优势木,而高密度林分中林木密集度分布与林木大小无关。[结论]红花尔基沙地樟子松天然林结构不合理,应选择病腐木及聚集分布的个体作为潜在调整对象,进行密度调整和结构优化。     

Characterization of a cinnamoyl derivative from broccoli (Brassica oleracea L. var. italica) florets

A new intact glucosinolate Cinnamoyl derivative [6'-O-trans-(4″- hydroxy cinnamoyl)-4-(methylsulphinyl) butyl glucosinolate] (A) has been isolated from Broccoli (Brassica oleracea L. var. italica) florets. The compound was isolated and characterized by LC, MS-ESI, FTIR, ¹H and ¹³C NMR as well as ¹H-¹H COSY, DEPT 135° spectrometric experiments.     

我国樟子松人工林天然更新研究进展

樟子松人工林天然更新问题的研究一直在进行,但人工林内幼苗数量过少及难以发育为幼树的问题一直难以彻底解决。文中综述了影响我国沙地樟子松人工林天然更新能力的因素,如光照、温度、水分、土壤环境、生物因子及人工营林技术等的研究概况。在研究过程中发现,樟子松人工林的遗传多样性与土壤微生物对樟子松人工林天然更新影响方面的研究尚少有报道,或可以为解决樟子松人工林天然更新问题提供更深入的思路。     

Comparison of terpenes in extracts from the resin and the bark of the resinous stem canker of Chamaecyparis obtusa and Thujopsis dolabrata var. hondae

A monoterpene and 15 diterpenes were isolated from the ethyl acetate extracts of the bark-glued resin from the resinous stem canker ofThujopsis dolabrata var.hondae Makino. A monoterpene (nezukone20) and 4 diterpenes (acetyl torulosol5, acetyl isocupressic acid8, acetyl abietinol11, and 7-methoxytotarol18) were characteristic constituents of the ethyl acetate extracts but were absent in then-hexane extracts from the resinous stem canker ofT. dolabrata var.hondae. These terpenes were first isolated fromT. dolabrata var.hondae andT. dolabrata. The available literature suggests that diterpene18 is a new compound. The resinous stem canker ofChamaecyparis obtusa Endlicher contained larger amounts of manool1,trans-communic acid6, and ferruginol12 and smaller amounts of isocupressic acid7 and abietinol10 than the resinous stem canker ofT. dolabrata var.hondae. The concentration of18 was less than 2% in the extracts, and the resinous stem canker ofC. obtusa lacked this compound. The resinous stem cankers ofC. obtusa andT. dolabrata var.hondae provided extracts 15.6 and 4.96 times, respectively, heavier than the healthy ones. Large differences in the ratios and compositions of terpenes were also observed between the resinous stem canker and the healthy trees. Terpenes isolated from the extracts contained many kinds of diterpene, especially the labdane-type diterpenes, in these diseased trees. These results suggest that the presence of labdane-type diterpenes is closely associated with the resinous stem canker or the causal fungi of this disease.Part of this report was presented at the 50th annual meeting of the Japan Wood Research Society, Kyoto, April 2000