露水草中20—羟基蜕皮甾酮的动态变化

采用ＨＰＬＣ技术对云南产鸭跖草科分属于３个族１０个属的１３种植物中的２０－羟基蜕皮甾酮进行分析,结果表明２０－羟基蜕皮甾酮广泛存在于已分析的该科植物中,但含量除露水草外均甚微,不具有化学分类学的意义。２０－羟基蜕皮甾酮在不同产地和不同生长季节的露水草中的含量变化,则与蝶类昆虫的活动显示有一定的相关性,从而提示２０－羟基蜕皮甾酮有可能是露水草与蝶类昆虫协同进化过程的具有化学生态学意义的代谢产物。     

平卧川牛漆甾酮对小菜蛾生长发育的影响(英文)

平卧川牛漆甾酮是紫背金盘中的主要植物蜕皮甾类 ,在筋骨草属植物中普遍存在。试验结果表明 ,用 5 0mg L平卧川牛漆甾酮处理小菜蛾卵 ,其孵化受到抑制 ,总孵化率为 94.7% ,显著低于对照的 1 0 0 %孵化。平卧川牛漆甾酮对幼虫具有弱的毒杀活性和良好的拒食活性 ,且与浓度相关。小菜蛾的生长发育也受平卧川牛漆甾酮的影响。用高于 5 0mg L浓度处理幼虫 ,其生长受抑及蛾的产卵量下降 ,而较低浓度处理则有利于幼虫生长 ,卵量提高。这种双重作用可能是由于平卧川牛漆甾酮的激素活性和拒食活性的作用结果 ,后者导致昆虫营养不良。处理幼虫后 ,试虫的化蛹和羽化受阻。小菜蛾对平卧川牛漆甾酮比 2 0 羟基蜕皮酮更敏感。     

20-羟基蜕皮甾酮处理后舞毒蛾外部形态和幼虫体壁超微结构的变化

为了探明20-羟基蜕皮甾酮对昆虫蜕皮过程中体壁的表皮层、 皮细胞及其细胞器的具体影响过程, 本研究利用透射电镜技术研究了20-羟基蜕皮甾酮对舞毒蛾Lymantria dispar （Linnaeus）5龄幼虫体壁超微结构的变化。结果表明, 用高浓度20-羟基蜕皮甾酮溶液浸过的白桦叶片饲喂幼虫, 处理6 h, 摄入约400 μg 20-羟基蜕皮甾酮后, 幼虫停止取食; 处理12 h时表皮细胞顶膜上的微绒毛减少, 在皮细胞与旧表皮之间形成蜕皮间隙, 旧头壳从幼虫头部脱离; 处理24 h时蜕皮间隙继续增大, 旧表皮与皮细胞进一步分离, 新表皮质层开始形成; 处理36 h时皮细胞顶膜形成较短的微绒毛, 胞质区域出现数量较多的电子疏松泡, 新表皮由上表皮、 外表皮及8层左右内表皮片层组成; 处理48 h时顶膜与内表皮界限模糊, 内表皮继续合成至16层左右; 72 h时细胞内出现大面积电子疏松泡, 内表皮合成至20层左右。 处理96 h时, 与对照组相比, 皮细胞细胞器较少, 核仁周围出现小部分空白区域, 胞质区域内含物减少; 虫体发黑缩小, 即将死亡; 内表皮层数仍旧保持20层左右。对照组幼虫6-96 h虫体活跃, 正常取食, 外部观察及透射电镜结果均未显现蜕皮现象; 表皮层由上表皮、 外表皮及内表皮组成; 皮细胞顶膜微绒毛密度高; 表皮细胞分泌活动旺盛, 胞质区域细胞界限明显, 内含物丰富; 细胞器典型而且活跃; 内表皮片层随时间不断增加至50层左右。结果提示, 外源20-羟基蜕皮甾酮能够导致舞毒蛾5龄幼虫的致死性蜕皮。     

沙打旺的化学成分

从沙打旺分离到１５个化合物,经光谱分析,理化实验及与已知化合物比较,鉴定了１１个化合物,其中两个为四环三萜类化合物,分别鉴定为（２０Ｒ,２４Ｓ）－３．１６－二羰基－６α,２５－二羟基－２０,２４－环氧－９．１９－环羊甾酮（９）和（２０Ｒ,２４Ｓ）－３,１６－二羰基－６α,２５－二羟基－２０,２４－环氧－９,１９－环－２３氧－羊毛甾酮（１０）。     

Changes in external morphology and integument ultrastructure of the 5th instar larvae of Lymantria dispar ( Lepidoptera: Lymantridae) treated by 20-hydroxyecdysone

为了探明20-羟基蜕皮甾酮对昆虫蜕皮过程中体壁的表皮层、皮细胞及其细胞器的具体影响过程,本研究利用透射电镜技术研究了20-羟基蜕皮甾酮对舞毒蛾Lymantria dispar( Linnaeus)5龄幼虫体壁超微结构的变化.结果表明,用高浓度20-羟基蜕皮甾酮溶液浸过的白桦叶片饲喂幼虫,处理6h,摄人约400 μg 20-羟基蜕皮甾酮后,幼虫停止取食;处理12h时表皮细胞顶膜上的微绒毛减少,在皮细胞与旧表皮之间形成蜕皮间隙,旧头壳从幼虫头部脱离;处理24 h时蜕皮间隙继续增大,旧表皮与皮细胞进一步分离,新表皮质层开始形成;处理36 h时皮细胞顶膜形成较短的微绒毛,胞质区域出现数量较多的电子疏松泡,新表皮由上表皮、外表皮及8层左右内表皮片层组成;处理48 h时顶膜与内表皮界限模糊,内表皮继续合成至16层左右;72h时细胞内出现大面积电子疏松泡,内表皮合成至20层左右.处理96 h时,与对照组相比,皮细胞细胞器较少,核仁周围出现小部分空白区域,胞质区域内含物减少;虫体发黑缩小,即将死亡;内表皮层数仍旧保持20层左右.对照组幼虫6 -96 h虫体活跃,正常取食,外部观察及透射电镜结果均未显现蜕皮现象;表皮层由上表皮、外表皮及内表皮组成;皮细胞顶膜微绒毛密度高;表皮细胞分泌活动旺盛,胞质区域细胞界限明显,内含物丰富;细胞器典型而且活跃;内表皮片层随时间不断增加至50层左右.结果提示,外源20-羟基蜕皮甾酮能够导致舞毒蛾5龄幼虫的致死性蜕皮.     

鳞翅目若干迁飞昆虫翅形特征分析(英文)

对鳞翅目蛾类 2科 42种和蝶类 7科 31种昆虫的翅形进行数值分析。结果表明 ,无论是蛾类还是蝶类 ,迁飞昆虫前翅形状均具有区别于非迁飞昆虫的共同特征 ,即前翅相对窄长 ,前缘较平直 ;然而与不同的翅振模式相适应 ,蛾类与蝶类迁飞昆虫的翅形特征又有所不同 :蛾类迁飞昆虫翅外侧较宽阔 ;蝶类迁飞昆虫翅外侧较窄小。     

棉铃虫蛹期血淋巴的蜕皮甾类

目前为止仅在少数几种昆虫中研究过蛹期的蜕皮激素。关于蜕皮甾类的性质分析,结果也颇不一致。本文采用放射免疫分析、薄层层析、高压液相色谱及质谱对棉铃虫Heliothis armigera蛹血淋巴内的蜕皮激素进行了研究。结果如下:1.物理-化学方法证明蛹血淋巴内存在二种蜕皮甾类:蜕皮酮和20-羟基蜕皮酮。2.蛹期蜕皮甾类滴度呈一宽峰,高峰出现在化蛹后的第5天(3435ng/ml)。3.在高峰时,蜕皮酮与20-羟基蜕皮酮的比例为1:3.57,说明20-羟基蜕皮酮是主要的蜕皮甾类。4.比较雌雄两性蛹的蜕皮甾类滴度,未见明显差异。研究表明在棉铃虫中影响成虫发育的主要激素是20-羟基蜕皮酮而不是蜕皮酮。     

20-羟基蜕皮甾酮对大鼠全脑缺血再灌注后海马神经元损伤和炎症反应的影响

目的:观察20-羟基蜕皮甾酮对全脑缺血再灌注后SD大鼠海马神经元和认知功能的保护作用,并探讨其相关机制。方法:采用四血管闭塞法建立SD大鼠全脑缺血再灌注模型,脑电图和脑组织Nissl染色评估模型的可靠性。将实验动物分为假手术组,缺血再灌注组和缺血再灌注+20-羟基蜕皮甾酮组。TUNEL染色观察海马神经元凋亡,Morris水迷宫实验评价大鼠的认知功能,酶联免疫法测定缺血再灌注后3-24小时大鼠血清中白细胞介素-1β(IL-1β)和肿瘤坏死因子α(TNF-α)的浓度。结果:全脑缺血再灌注后大鼠海马神经元凋亡率从4.50±1.90%上升至72.90±8.40%(p0.01),给予20和40 mg/kg 20-羟基蜕皮甾酮干预,大鼠海马神经元凋亡率分别下降至51.40±8.60%(p0.05)和42.70±6.80%(p0.01)。与假手术组相比,全脑缺血再灌注后大鼠在Morris水迷宫定位航行试验中逃避潜伏期明显延长(p0.01),在空间探索试验中目标象限停留时间和穿越目标象限次数明显减少(p0.01),而20-羟基蜕皮甾酮显著抑制上述变化,改善大鼠的认知功能。缺血再灌注后3-24小时,大鼠血清中IL-1β和TNFα浓度较假手术组显著升高,20-羟基蜕皮甾酮能抑制上述各时间点大鼠血清中IL-1β和TNFα浓度的升高。结论:20-羟基蜕皮甾酮对全脑缺血再灌注后大鼠海马神经元和认知功能有显著保护作用,抑制缺血再灌注后的炎症反应是其保护机制之一。     

露水草家化栽培研究简报

露水草(Cyanotis arachnoidea C.B.Clarke),是近年发现提取昆虫蜕皮激素的优良原料植物,是迄今发现的植物中含β-蜕皮激素最多的种之一。它的干燥全草中昆虫蜕皮激素的含量达1.2％,地下部分达干重的2.9％。把昆虫激素用于养蚕,是近代科学养蚕的重要技术,可使家蚕上簇齐一,减少用桑,节约劳力。因此对它进行引种栽培方面的研究,为工业生产昆虫蜕皮激素提供优质原料,具有较大的现实意义。1978—1982年我们对这一新的资源植物,进行了家化栽培研究,现报道如下。     

浙江天童国家森林公园蝶类昆虫多样性研究

为探明浙江天童国家森林公园蝶类昆虫本底资料,提供该地区后续开发利用、进一步保护蝶类资源相应的理论依据,开展本项目研究。对已有和补充采集标本整理鉴定的基础上,结合国内外已公开发表的资料,建立浙江天童国家森林公园鳞翅目Lepidoptera蝶类昆虫数据库。共记录鳞翅目蝶类昆虫10科58属82种,其中14种(隶属于6科14属)为本研究在该地区新纪录种类。其寄主涉及32科81属植物,不同科的蝶类对寄主植物的选择具有一定专化性。蝶类区系类型组成以东洋+古北共有种类为主,共47种,东洋种32种,古北种3种,分别占总数的57.32%,39.02%和3.66%,表明古北界与东洋界的蝶类物种在该地区有广泛的交流。     

Photosynthetic Characteristics of Cyanotis arachnoidea and Their Ecological Significance

The photosynthetic characteristics of Cyanotis arachnoidea Clarke was studied through shading experimentation in pot culture. The effect of different shade level on the photosynthetic rate, the characteristics of photosynthetic organs and the bio-production of C. arachnoidea were compared. The main results are as follows: 1. C. arachnoidea was a C3 type solaphilic and shade tolerant herb. Its light saturation point was about 650 μmol · m-2 · s-1 and the light compensation point was about 17 μmol · m-2 · s-1. Its CO2 compensation point was about 130× 10-6. 2. The maximum net photosynthetic rate of C. arachnoidea was about 12.45 μmol · m-2 · s-1. The diurnal variation regularity of its photosynthetic rates exhibited a double peak curve in which the main peak appeared during 11: 00～12: 00, and the second around 15: 00. 3. 20%～50% shading favored the growth of C. arachnoidea. In comparison with those with control treatment, the content of chlorophyll b with 20%～50% of shading treatment increased by 47 %～83%, and the photosynthetic production increased by 12 %～18 % as a result of increase in the net photosynthetic rate (relative photosynthetic rate).     

Phytoecdysteroids from the juice of Serratula coronata L. (Asteraceae).

Seven phytoecdysteroids have been isolated from Serratula coronata L. One of them is a new phytoecdysteroid, 3-epi-20-hydroxyecdysone. Two further ecdysteroids, 20-hydroxyecdysone 22-acetate and taxisterone, are isolated from this species for the first time in addition to the typical S. coronata ecdysteroids, 20-hydroxyecdysone, ecdysone, ajugasterone C and polypodine B. The juice squeezed from aerial parts of fresh plants of S. coronata was extracted with ethyl acetate. The ecdysteroids were isolated by a combination of chromatographic techniques (mainly HPLC) and identified by 1D and 2D (1)H and (13)C NMR experiments and mass-spectrometry. The biological activities of 3-epi-20-hydroxyecdysone (EC(50)=1.6 x 10(-7) M), taxisterone (EC(50)=9.5 x 10(-8) M) and ajugasterone C (EC(50)=6.2 x 10(-8) M) have been determined in the Drosophila melanogaster B(II) bioassay for ecdysteroid agonist activity.     

The Effects of Ingested 20-Hydroxyecdysone on the Larvae of Aglais urticae,Inachis io,Cynthia cardui (Lepidoptera: Nymphalidae) and Tyria jacobaeae (Lepidoptera: Arctiidae)

A comparative survey was carried out to investigate the effects, distribution and metabolism of ingested 20-hydroxyecdysone in four species of lepidopteran larvae in relation to the phytoecdysteroid content of the insect's host plants. Analysis of the leaves of the host plants of each of the species revealed a strong relationship between the levels of phytoecdysteroids and the relative tolerance of the larvae to ingested 20-hydroxyecdysone. Monophagous or oligophagous species (Aglais urticae, Inachis io) feeding on ecdysteroid-negative host plants were either deterred from feeding or showed marked abnormalities in growth and development after incorporation of 20-hydroxyecdysone in their diets. Oligophagous or polyphagous species (Tyria jacobaeae, Cynthia cardui) which feed on host plants from families which are known to contain phytoecdysteroid-positive species, were able to tolerate low levels of 20-hydroxyecdysone in their diets, but exhibited developmental defects at high concentrations. These species were termed semi-tolerant. In each of the species, ingested [3H]20-hydroxyecdysone appeared to follow the same fate as injected [3H]20-hydroxyecdysone. The data are compared to those obtained in previous studies, where truly polyphagous species were shown to tolerate very high concentrations of 20-hydroxyecdysone in their diets by the production of ecdysteroid 22-fatty acyl esters. Copyright 1997 Elsevier Science Ltd. All rights reserved     

Phylogeographic patterns in Hypochaeris section Hypochaeris (Asteraceae, Lactuceae) of the western Mediterranean

Aim  To analyse phylogeographic patterns in the four species of Hypochaeris sect. Hypochaeris , evaluating possible areas of origin and the microevolutionary processes that have shaped their morphology, genetics and distribution.
Location  Western Mediterranean area.
Methods  We applied amplified fragment length polymorphism (AFLP) markers to a total of 494 individuals belonging to 82 populations of Hypochaeris arachnoidea , H. glabra , H. radicata and H. salzmanniana to determine population structure.
Results  Populations with the largest proportion of private and rare AFLP fragments were found in Morocco. This region was consequently inferred to be the ancestral area for H. arachnoidea , H. glabra , H. radicata and H. salzmanniana . The Guadalquivir River (southern Spain) was inferred to be an effective dispersal barrier for H. glabra and H. radicata. The Strait of Gibraltar was inferred to be a somewhat weaker barrier than the Guadalquivir River for H. radicata and a much weaker barrier for H. glabra . The main barrier for H. salzmanniana coincides with the extension of the Rif Mountains to the Atlantic coast in Morocco, and the Strait of Gibraltar is a much weaker barrier for this species. Hypochaeris arachnoidea appears to have originated in the Atlas Mountains.
Main conclusions  The highest levels of genetic variation in La Mamora forest ( H. glabra and H. salzmanniana ) or the adjacent central Middle Atlas ( H. arachnoidea and H. radicata ) in Morocco suggest that these areas were a centre of origin of Hypochaeris sect. Hypochaeris . All three potential barriers – the Guadalquivir River, the Strait of Gibraltar, and the Rif Mountains – have been important in shaping genetic diversity in species of section Hypochaeris .     

Mechanism of formation of the A/B cis ring junction of ecdysteroids in Polypodium vulgare.

1. The fates of the alpha-, 4 alpha- and 4 beta-hydrogen atoms of cholesterol during formation of the A/B cis ring junction of ecdysteroids was investigated by administration of [4-14C, 3 alpha-3H], [4-14C, 4 alpha-3H]- and [4-14C, 4 beta-3H]cholesterol species to the fern, Polypodium vulgare, and isolation of the 20-hydroxyecdysone formed in each case. 2. The 3H was retained in the ecdysteroid formed from each substrate. 3. Location of the 3H in the 20-hydroxyecdysone indicated that migration of 3H from the 3 alpha- and 4 beta-positions to C-4 and C-5, respectively, had occurred, whereas the 4 alpha-3H atom was retained at C-4. 4. A possible mechanism for the formation of the A/B cis ring junction of ecdysteroids in P. vulgare is presented.     

Induction of malate dehydrogenase and acetylcholinesterase by 20-hydroxyecdysone and heat shock in Drosophila ovaries

The molting hormone 20-hydroxyecdysone induces de novo synthesis of cytoplasmic malate dehydrogenase (cMDH) in Drosophila ovaries, but not mitochondrial MDH (mMDH). A second enzyme, acetylcholinesterase (AChE), was found to be heat shock inducible. It is known that MDH and AChE are, respectively, heat shock and 20-hydroxyecdysone inducible (see Introduction). Now it is also known that these enzymes are under dual regulation, with 20-hydroxyecdysone and heat shock being two stimuli which act either separately or in combination, to increase the specific activity of these enzymes. The response to 20-hydroxyecdysone and/or heat shock was found to occur in seven additional D. melanogaster sibling species. In this case, hormone and heat shock maximize the interspecific variability, something which could be acted by natural selection to establish physiological adaptations.     

Ecdysone 20-monooxygenase activity in Drosophila virilis strains varying in ecdysteroid response to heat stress

The effect of various duration of heat stress (38 degrees C) on the activity of ecdysone 20-monooxygenase converting ecdysone into 20-hydroxyecdysone has been studied in D. virilis of wild type and mutant strain females, which differ by the mode of heat stress response of ecdysone and 20-hydroxyecdysone. We are the first to show that heat stress induces activity of ecdysone 20-monooxygenase in Drosophila females and enzyme activity correlates with the level of 20-hydroxyecdysone.     

The effects of 20-hydroxyecdysone on the differentiationin vitro of cells from the eye imaginal disc fromDrosophila melanogaster

We have examined the effects of the insect ecdysteroid, 20-hydroxyecdysone, on the differentiation of neuronal and non-neuronal elements in the developing adult visual system, usingin vitro methods inDrosophila. We examined the differentiation of early neuronal markers in the presence and absence of 1 μg/ml 20-hydroxyecdysone. Immunoreactivity to 22C10, a marker of an early neuronal antigen, as well as to the photoreceptor-specific antibody 24B10, suggests that differentiation of neuronal and photoreceptor antigens does not require 20-hydroxyecdysone. In eye-discs cultured from animals 5 hours after the white prepupa (P+5), ommochrome pigmentation first appeared after 2 days in 1 μg/ml 20-hydroxyecdysone, but cultures lacked pigmentation without 20-hydroxyecdysone. Our culture conditions failed to support the formation of the second screening pigment, drosopterins, even with 20-hydroxyecdysone. Eye discs from P+5 also formed lenses and interommatidial bristles in culture when 20-hydroxyecdysone was added but not in cultures devoid of the hormone. The differentiation of synaptotagmin and the elongation of extending photoreceptor neurites from eye disc fragments both occur in the absence of 20-hydroxyecdysone in cultures, but adding the hormone increased average neurite length. The threshold for enhanced neurite length was less than 125 ng/ml 20-hydroxyecdysone. Eye-disc cultures also developed immunoreactivity to histamine, the photoreceptor transmitter, from synthesis not re-uptake, in both the presence and in the absence of 20-hydroxyecdysone. These findings suggest that photoreceptor axons may be able to release transmitterin vivo both when they grow into the optic lobe and during the subsequent events in synapse formation.     

Interendocrine control by 20-hydroxyecdysone of the corpora allata of Manduca sexta

A positive interendocrine control by 20-hydroxyecdysone regulates the corpora allata (CA) and thus the hemolymph titers of the juvenile hormones (JHs) during the fourth and fifth larval stadia of the tobacco hornworm, Manduca sexta. The effect of 20-hydroxyecdysone is exerted via the brain-corpora cardiaca (Br-CC), and the kinetics of stimulation by the hormone suggest that its control of the CA is indirect, possibly involving neural effectors. Co-incubation of precommitment CA with Br-CC resulted in a stimulation of the synthesis of JH I acid equivalent to that achieved by the incubation of brain-corpora cardiaca-corpora allata (Br-CC-CA) with 20-hydroxyedcysone, suggesting that a diffusible regulatory factor may be involved in the stimulation. During pharate pupal development, a close temporal relationship exists between the drop in the ecdysteroid titer and declines in CA activity and the JH hemolymph titer, suggesting that 20-hydroxyecdysone may now inhbit the CA. Incubations of day 6 Br-CC-CA with physiological concentrations of 20-hydroxyecdysone resulted in an apparent dose-dependent inhibition of CA activity. Thus in Manduca, 20-hydroxyecdysone may exert varying stage-specific interendocrine effects on the CA which direct the postembryonic development of this species.     

露水草毛状根的诱导和培养

发根农杆菌（Ａｇｒｏｂａｃｔｅｒｉｕｍｒｈｉｚｏｇ６ｎｅｓ）ｐＲｉ１５８３４菌株感染露水草（Ｃｙａｎｏｔｉｓａｒａｃｈｎｏｉｄｅａ）的茎和根的外植体及小苗,均诱导出毛状根。毛状很能在不含激素的ＭＳ培养基上生长。用纸电泳和高效薄层层析法,在毛状根中检测到甘露碱。用Ｓｏｕｔｈｅｒｎ印迹杂交,在植物ＤＮＡ中检测到ｒｏｌｅ基因。表明Ｒｉ质粒的Ｔ—ＤＮＡ部分已转移到毛状根细胞的ＤＮＡ中。该毛状根能产生β－蜕皮激素。露水草毛状根的诱导培养在单子叶植物中是一个成功的先例。