十种虫生真菌液体培养芽生孢子的形态发生

芽生孢子是许多真菌通过出芽方式产生的无性孢子,在液体发酵中主要以这种形式大量繁殖。真菌杀虫剂活性成分分生孢子的大量生产使用液体发酵得到的芽生孢子作为接种物,另外,它也是当今虫生真菌遗传转化的重要受体,因而研究虫生真菌芽生孢子的形态和发生特点具有重要意义。本研究分别对液体发酵中的球孢白僵菌、粉棒束孢、蝉棒束孢、环链棒束孢、玫烟色棒束孢、细脚棒束孢、斜链棒束孢、金龟子绿僵菌、蝗绿僵菌和蜡蚧霉等10种常见虫生真菌芽生孢子的形成过程进行显微观察,了解其分生孢子产生过程的异同。结果表明,芽生孢子的产生方式有两种类型：1）蝉棒束孢在其整个生活史中以菌丝生长为主,芽生孢子产生的数量很少。2）其他各种真菌芽生孢子产生方式相似,在菌丝体形成后就开始大量以菌丝出芽或缢缩产生芽生孢子,接着还可通过芽生孢子的出芽或缢缩断裂产生新的芽生孢子。芽生孢子的产生分为3个时期：初期先由菌丝形成芽生孢子;指数期芽生孢子大量增殖,菌丝和芽生孢子都可产生芽生孢子;后期以芽生孢子产新芽生孢子为主要方式。     

湖南白檀居群形态多样性及与环境的相关性

为了解白檀在湖南地区不同环境的适应性及遗传变化程度,对湖南地区6个白檀天然居群的189个个体的11个表型性状进行形态遗传多样性分析。结果表明,11个表型性状均具有丰富的遗传多样性(H'=1.389),居群间的平均方差分量为52.60%,居群内的平均方差分量为37.03%,说明在白檀形态性状多样性分布上是居群间多样性程度大于居群内,即居群间的形态变异是白檀形态变异的主要来源。聚类分析结果表明,6个居群的表型性状并没有严格依地理距离聚类,主成分与相关分析结果显示在11个表型中树型因子是主要形态变异因子,年降雨量与树型因子呈显著正相关,其他表型变异受遗传的影响可能大于受环境的影响。     

一株感染杨尺蠖蛹的病原真菌的分离及鉴定

【目的】从山西省洪洞县的杨树丰产林中采集到几头自然染菌的杨尺蠖Apocheima cinerarius越冬蛹虫尸,经分离纯化得到一株病原真菌,定名为YHT01,确定该菌株的分类地位。【方法】用该菌株孢子悬浮液(1.0×108 conidia/m L)感染杨尺蠖蛹进行回接杀虫实验,扫描电镜观察菌株形态学特征,ITS序列测定并构建系统发育树对YHT01菌株进行分子鉴定。【结果】经YHT01菌株感染之后,杨尺蠖蛹节间处长出白色菌丝。该菌株在PDA培养基上的菌落呈椭圆形,白色,绒毛状,菌落背面呈橙黄色;扫描电镜观察结果显示,该菌株孢梗束呈扫帚状,分生孢子小,卵形至椭圆形,壁光滑,大小为(2.1-3.4)μm×(1.1-1.8)μm。该菌株的r DNA-ITS序列与Gen Bank中粉棒束孢Isaria farinosa(Holm:Fr.)Fr.(AB233337)的序列相似性达99%,在系统发育树的同一分支上。【结论】YHT01是杨尺蠖蛹的一株病原真菌,粉棒束孢Isaria farinosa。     

小叶锦鸡儿天然居群叶形态性状变异研究

为揭示小叶锦鸡儿(Caragana microphylla)天然居群叶形态性状的变异规律及其生态适应性特征,该研究以10个小叶锦鸡儿天然居群为对象,通过多重比较、巢式方差分析、相关性分析、聚类分析和主成分分析等方法,对7个叶形态性状进行分析。结果表明：(1)小叶锦鸡儿叶形态性状在居群内和居群间均存在极显著差异(P < 0.01),平均变异系数为10.13%,不同性状的变异幅度为6.23%~12.78%;平均叶形态性状的表型分化系数为43.62%,居群内变异(30.09%)大于居群间变异(24.91%),说明居群内是其叶形态性状变异的主要来源。(2)相关性分析表明,环境因子对小叶锦鸡儿的叶形态性状变异有很大的影响,在地理空间上主要呈现出沿海拔梯度的变异模式;主成分分析的结果显示,小叶宽、叶柄宽和叶柄长对小叶锦鸡儿叶形态变异起主导作用;利用欧式距离对小叶锦鸡儿居群进行UPGMA聚类分析结果显示,基于叶形态性状和环境因子可分别将小叶锦鸡儿10个居群分为3类和2类,Mantel检验结果表明,小叶锦鸡儿的叶形态性状变异不存在地理连续性。研究结果为小叶锦鸡儿的适应性进化和开发利用提供了理论依据。     

云南虫生真菌粉棒束孢遗传分化研究

对云南粉棒束孢8个当地居群和蝙蝠蛾拟青霉2个当地居群进行ITS测序和RAPD扩增分析,结合Gen Bank中相关序列,对粉棒束孢开展遗传多样性、居群遗传结构及其种内分化研究。共获得大范围内地理距离远的6个居群共97条粉棒束孢ITS序列,共有33种单倍型,单倍型多样性Hd=0.546和总核苷酸多样性Pi=0.00276,显示粉棒束孢在物种水平上遗传多样性较低。云南粉棒束孢共37条序列,有14种单倍型(10种为云南特有),具有较高单倍型多样性和核苷酸多样性(Hd=0.659,Pi=0.00274);单倍型聚类和网状分支分析表明云南粉棒束孢单倍型类型丰富,遗传多样性高,暗示云南为粉棒束孢多样性分布中心之一。ITS序列分析表明,云南当地居群间遗传分化系数Fst=51.95%;RAPD分析表明,居群间遗传分化系数Gst=0.5547,基因流Nm=0.4014;说明云南当地居群粉棒束孢分化剧烈。居群遗传距离与地理距离相关性研究表明,粉棒束孢居群遗传距离与地理距离无明显相关。中性检验和失配分析表明粉棒束孢经历过近期居群扩张。结合单倍型聚类和网状分支分析,表明Hap 19为扩张建群单倍型,但原始祖先单倍型(Hap 1)依然是粉棒束孢居群中最优势单倍型(频率为48.45%),表明粉棒束孢并不存在明显的因地理原因造成的生殖隔离。值得重视的是,通过ITS单倍型和RAPD分析,支持将蝙蝠蛾拟青霉作为粉棒束孢异名处理。     

胶孢炭疽菌生防放线菌SD-29的鉴定及抗菌活性评价

【背景】胶孢炭疽菌是引起橡胶炭疽病的一种重要病原菌,可导致橡胶树产胶量下降。【目的】从山东青岛一农田土壤中分离出一株胶孢炭疽菌生防放线菌SD-29,并对其进行鉴定及抗菌活性评价。【方法】采用对峙生长法及菌丝生长速率法对菌株SD-29的拮抗活性进行鉴定;利用乙酸乙酯萃取法提取菌株SD-29发酵液粗提物并进行活性评价;根据菌株SD-29的形态特征、生理生化及16S rRNA基因序列进行鉴定。【结果】菌株SD-29对胶孢炭疽菌具有较强的抑制活性,皿内抑制活性达到82.6%。发酵液粗提物对菌丝生长的EC50为13.6μg/mL,100μg/mL的粗提物对胶孢炭疽菌孢子萌发抑制率达到63.16%,其对感炭疽病橡胶叶片的防治效果达到48.96%。根据该菌的形态特征、生理生化特征及16S rRNA基因序列分析鉴定菌株SD-29为Streptomyces yatensis。【结论】菌株SD-29对胶孢炭疽菌有较强的防治效果,具有潜在的应用价值。     

爪哇棒束孢侵染美国白蛾过程及抗氧化酶活性变化

[目的]爪哇棒束孢Isaria javanica是一种重要的虫生真菌,已广泛应用于多种害虫的防治.本研究从美国白蛾Hyphantria cunea僵虫尸体中分离出对美国白蛾具有致病作用的爪哇棒束孢BE01菌株,为了解该菌株对美国白蛾的侵染过程,并通过研究宿主体内抗氧化酶系对爪哇棒束孢BE01菌株入侵的响应,评价该菌的杀虫作用,为爪哇棒束孢BE01防治美国白蛾提供依据.[方法]采用扫描电镜技术,观察爪哇棒束孢BE01分生孢子侵染的美国白蛾3龄幼虫,并测定美国白蛾接菌后体内超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)含量的变化.[结果]爪哇棒束孢对美国白蛾侵染过程包括:分生孢子粘附在美国白蛾的角质层上,接种后6h孢子开始萌发,24h产生附着胞,48 h菌丝在体表继续生长,96h菌丝从虫体内长出并产分身孢子,120 h菌丝覆盖整个虫体.在受到爪哇棒束孢BE01侵染后,美国白蛾幼虫体内的3种抗氧化酶SOD、POD、CAT酶活性明显升高,并在侵染48 h时达到峰值.随着侵染时间的增加,48 h后3种酶的活性开始下降, 60 h后酶活性均低于对照组.[结论]爪哇棒束孢BE01孢子活力高、萌发速率快,导致美国白蛾体内的保护酶系难以发挥保护作用.爪哇棒束孢BE01菌株能够快速侵染美国白蛾幼虫,具有开发成新的高效生物防治菌株的潜能.     

不同培养时间下蝉棒束孢菌丝体的代谢组研究

利用LC-MS对不同培养时间蝉棒束孢菌丝体代谢物进行检测,结合多变量统计分析的主成分分析(PCA)和正交偏最小二乘判别分析(OPLS-DA)鉴定差异化合物。PCA和OPLS-DA表明蝉棒束孢菌丝体中的代谢物随培养时间不同有显著变化。阴、阳离子模式下我们都可以把蝉棒束孢代谢物变化分为前期、中期和后期3个时期。培养前期产生较多的细胞松弛素E、肌醇、γ-氨基丁酸、乙酰肉碱、亚油酸等与侵染宿主、早期信号传导和氧化供能有关的物质;中期产生较多的甘露醇、海藻糖、亚麻酸、柠檬酸、丙酮酸、7,8-二羟基油酸等与抗逆和氧化供能有关的物质,此外还有两种抗生素红酵母红素、δ-氨基-γ-酮戊酸达到了最大值;培养后期产生较多的脯氨酸、2,4-二氨基丁酸、3′-deoxy-hanasanagin、ulvaline、赤霉素、烟酰胺等与抗逆、抗氧化、产孢、细胞保护有关的物质。首次发现烟酰胺可能与蝉棒束孢从营养生长转化为生殖生长及产孢有关;同时赤霉素、ulvaline和δ-氨基-γ-酮戊酸可能与蝉棒束孢产孢或孢子成熟有关,尤其值得注意的是蝉棒束孢在培养后期高速合成和大量积累赤霉素,值得深入研究。根据蝉棒束孢的代谢特点,作为药品及保健品开发蝉棒束孢菌丝产品时,培养时间宜在7–11d。     

稻曲病菌厚垣孢子脂肪酸组成的研究

【目的】为探明细胞壁和细胞内脂肪酸成分及含量与细胞抗逆性的关系,【方法】采用酸热法、索氏提取法、有机溶剂法对稻曲病菌的厚垣孢子壁进行脂肪酸提取,并采用气相色谱检测其脂肪酸的组成和含量。【结果】采用酸热法提取脂肪酸效果最好,以该方法提取测定稻曲病菌黄色、黄绿色、黑色厚垣孢子壁饱和脂肪酸相对含量分别为26.92%、17.23%、23.71%,其不饱和脂肪酸相对含量分别为60.46%、61.52%、70.64%;厚垣孢子总(沉淀孢子壁和上清液)饱和脂肪酸相对含量分别为28.87%、21.00%、24.04%,厚垣孢子总不饱和脂肪酸相对含量分别为55.43%、55.87%、63.89%。硬脂酸在厚垣孢子壁中的含量:黄色>黄绿色>黑色;不饱和脂肪酸中顺式-5,8,11,14,17二十碳五烯酸(EPA)在厚垣孢子壁的含量:黑色>黄绿色>黄色。【结论】在3种颜色厚垣孢子中,黑色休眠型厚垣孢子在孢子壁、总不饱和脂肪酸含量均最高,表明不饱和脂肪酸的含量提高,有利于厚垣孢子的休眠越冬。     

贝莱斯芽孢杆菌GDND-2对烟草镰孢根腐病菌生长发育的影响

【背景】尖孢镰孢(Fusariumoxysporum)引起的烟草根腐病在世界烟区普遍发生,严重影响烟草产量和质量,化学农药无法有效防治病害,利用生防菌防治该病成为研究热点。【目的】明确贝莱斯芽孢杆菌(Bacillusvelenzensis)GDND-2对尖孢镰孢生长发育的抑制作用。【方法】制备含10%和20%GDND-2发酵滤液的PDA培养基,涂在玻片上,接种尖孢镰孢分生孢子,观察滤液对尖孢镰孢孢子萌发、菌丝生长、孢子形成和色素产生的影响,应用扫描和透射电镜观察菌体超微结构的变化。【结果】贝莱斯芽孢杆菌GDND-2发酵滤液延迟孢子萌发2h以上,造成芽管膨大畸形,促使菌丝提早分枝,抑制菌丝延伸,使菌丝产生畸形球状结构,10%和20%发酵滤液对菌丝生长抑制率达53.41%和61.58%。滤液延迟病菌产孢,显著抑制产孢量,影响孢子形态,刺激病菌产生色素。10%和20%发酵滤液延迟产孢20h和28h,对产孢量的抑制率为52.11%和78.85%,滤液促使病菌形成小型分生孢子。观察尖孢镰孢的超微结构,部分菌丝膨大、畸形,细胞壁变薄,细胞膜消失,细胞质渗出,胞内呈空腔结构;部分菌丝严重皱缩、扭曲、...     

The fine structure of conidial development in the genus Torula. I. T. herbarum (Pers.) Link ex S. F. Gray and T. herbarum f. quaternella Sacc.

Conidiogenesis in Torula herbarum and T. herbarum f. quaternella was observed by scanning and transmission electron microscopy. Conidia of the former were shown to be made up of three equally sized cells capped by a distinctive, and easily recognizable, conidiogenous cell. Conidiogenous cells also arose terminally on erect hyphae and on prostrate hyphae. The single-layered conidial cell walls were differentiated into an inner hyaline zone and an outer electron-dense zone formed by the deposition of melanin. Conidiogenous cells lacked melanin at the apex and, before conidiation, the lateral walls were strengthened by a further deposition of melanin. The apex bulged outwards and was modified into a new multicelled conidium bearing another apical conidiogenous cell. Continued development of new conidia resulted in an acropetal chain which became disarticulated after cytolysis within the conidiogenous cell. The relative distinctions between holoblastic and enteroblastic development are discussed and it is concluded that the conidia should be referred to as blastoconidia.     

Studies on the phylogenesis of pathogenic ‘black yeasts’

This paper deals mainly with the conidium ontogenesis and phylogenesis of black yeasts such as E. jeanselmei, E. gougerotii, E. dermatitidis and E. spinifera. The conidium ontogenesis of E. jeanselmei, E. gougerotii and E. dermatitidis was almost the same. One to five annellated tips were observed through a scanning microscope at the apices of conidiogenous cells, which were bottle- or jar-shaped. Annellations on the tips looked like fringes and the conidiogenous cells of these three species were annellides. Annellated projections occurred on hyphae and annelloconidia were also produced from them. Occasionally, secondary annellides occurred from primary ones. They looked like moniliform hyphae. Daughter conidia sometimes budded directly from mother cells. The shapes and sizes of the conidia of these species were very similar to each other. The conidium ontogenesis of E. spinifera was annellidic as well. However, a single annellated tip usually occurred on an annellide. The annellated tips of the fungus were long and more than 20 annellations were observed on their walls. The conidiogenesis of the four species of Exophiala is only annellidic. There were no differences in the biological examinations except KNO3 assimilation among these four species. The growth of E. jeanselmei and E. gougerotii was poor at 37 degrees C. The GC contents of E. jeanselmei 1171, E. gougerotii B-1800, E. dermatitidis MM-7 and E. spinifera DU-3342 were 54.6, 54.6, 56.6 and 59.7%, respectively.     

人工培养柞蚕蝉花不同部位的代谢组差异

[背景]蝉花是传统中药材,前期研究发现柞蚕培养蝉花与野生蝉花成分类似,因此有替代野生蝉花利用的潜力.[目的]揭示柞蚕培养蝉花各部分的代谢组差异,为合理利用人工培养柞蚕蝉花提供理论依据.[方法]用基于HPLC-DAD-TOF/MS代谢组学方法,比较人工培养柞蚕蝉花的虫体、体表菌丝和孢梗束的代谢组差异.[结果]人工培养柞蚕...     

Molecules and morphology in concert: tests of some hypotheses in arctic Potentilla (Rosaceae)

We developed a combined molecular and morphological approach to unravel complex variation at low taxonomic levels, exemplified by some arctic members of Potentilla. Twenty-one populations from Svalbard were analyzed for random amplified polymorphic DNAs (RAPDs) and 64 morphological characters to test the hypotheses that (1) the P. nivea complex (section Niveae) consists of three taxa (P. chamissonis, P. insularis, and P. nivea), (2) three "eco-morphotypes" in P. pulchella (section Multifidae) should be considered different taxa, and (3) P. insularis originated as an intersectional hybrid (Niveae × Multifidae). Twenty-two RAPD multilocus phenotypes were observed in the 136 plants analyzed based on 35 markers. Three fairly distinct groups of RAPD phenotypes were identified in the P. nivea complex based on multivariate analyses and an analysis of molecular variance (AMOVA; 77.6% among-group variation). The variation within the P. nivea complex was more or less continuous in multivariate analyses of the morphological data. We identified, however, several individual morphological characters that separated unambiguously among the three groups of RAPD phenotypes, revealing that these groups correspond to the previously hypothesized taxa. Many identical RAPD multilocus phenotypes were observed in the "eco-morphotypes" of P. pulchella, suggesting that its conspicuous morphological variation is caused by plasticity or by genetic variation at a small number of loci. The hypothesis of the hybrid origin of P. insularis was not supported by the RAPD data. Overall, very little RAPD variation was observed within populations of the four taxa (2.1-16.7% in AMOVA analyses; average genotypic diversity, D, was 0.10-0.30). We conclude that detailed, concerted analysis of molecules and morphology is a powerful tool in low-level taxonomy.     

蛹虫草(Cordycepsmilitaris)无性型的多型现象

在萨氏琼脂和PDA上,蛹虫草Cordycepsmilitaris（Vuill）Fr.无性型,蛹草拟青霉Paecilomycesmilitaris(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,大多数单孢子株属此类群。（2）属此类群的单孢子株,在PDA上可形成亮褐色至橙黄色的角变,它们典型的产孢结构为瓶梗轮生,分生孢子常聚集成头状的轮枝孢型（Verticillium－type）。在查氏培养基上则相反,拟青霉型的产孢结构占优势。单孢子株Cm-71是这一类群的代表。（3）单孢子株Cm－42是在所观察单孢子株中形态特征独特的菌株。在PDA上菌丝生长纤细,蛛网状,产孢结构轮枝孢型。蛹草拟青霉对氯酸钾（KClO3）不敏感,在浓度为6％（W／V）的KMM培养基上,23～24℃14天菌落的直径仅1～2cm,无气生菌丝生长。各单孢子株形成的不利用硝酸盐突变株（nit突变株）的途径和形态特征,也可分成类似于粉被虫草CordycepspruinosaPeteh不利用硝酸盐突变株（nit突变株）的三种类型。     

蛹虫草(Cordyceps militaris)无性型的多型现象*

在萨氏琼脂和PDA上,蛹虫草Cordycepsmilitaris（Vuill）Fr.无性型,蛹草拟青霉Paecilomycesmilitaris(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,大多数单孢子株属此类群。（2）属此类群的单孢子株,在PDA上可形成亮褐色至橙黄色的角变,它们典型的产孢结构为瓶梗轮生,分生孢子常聚集成头状的轮枝孢型（Verticillium－type）。在查氏培养基上则相反,拟青霉型的产孢结构占优势。单孢子株Cm-71是这一类群的代表。（3）单孢子株Cm－42是在所观察单孢子株中形态特征独特的菌株。在PDA上菌丝生长纤细,蛛网状,产孢结构轮枝孢型。蛹草拟青霉对氯酸钾（KClO3）不敏感,在浓度为6％（W／V）的KMM培养基上,23～24℃14天菌落的直径仅1～2cm,无气生菌丝生长。各单孢子株形成的不利用硝酸盐突变株（nit突变株）的途径和形态特征,也可分成类似于粉被虫草CordycepspruinosaPeteh不利用硝酸盐突变株（nit突变株）的三种类型。     

Morphological and allozymic variation in Hynobius boulengeri and H. stejnegeri (Amphibia: Urodela: Hynobiidae)

We studied morphological and allozymic variation in populations of Japanese salamanders, Hynobius boulengeri and H. stejnegeri. Adult H. boulengeri showed sexual dimorphism, and juveniles differed greatly from adults in many morphological characters. From the results of multivariate analyses of morphological characters, the populations were divided into four groups: (I) H. boulengeri from Honshu, (II) H. boulengeri from Shikoku, (III) H. boulengeri from the Sobo-Katamuki Mountains of Kyushu and H. stejnegeri, and (IV) H. boulengeri from the Amakusa Islands and the Osumi Peninsula. Phenotypic relationships among the four groups were identical to relationships clarified by allozymic analyses, except for group IV, which was included in group III in the allozyme tree. Some morphometric characters were significantly correlated with environmental variables. We consider H. stejnegeri to be a valid species based on its unique color pattern, morphometric characters, and allelic composition, even though it was nested within group III of H. boulengeri by both morphological and allozymic analyses. We propose that group I from Honshu and group II from Shikoku should be treated as H. boulengeri sensu stricto and H. hirosei, respectively. Resolving the taxonomic status of the remaining populations of groups III and IV from Kyushu requires further study.     

哈氏木霉分生孢子发育的超微结构和细胞化学

通过电子显微镜和细胞化学标记研究了哈氏木霉分生孢子发育的超微结构和细胞化学。分生孢子发育的超微结构研究表明,分生孢子壁的发育是有个由薄而光滑到厚而有疣的过程;期间脂肪体在分生孢子和产孢细胞中不断累积,最后脂肪体沿着内壁排列成一层。免疫金标记结果显示,幼嫩的分生孢子壁中缺乏几丁质和纤维素,只有在成熟的分生孢子壁中含有几丁质;出乎意料的是在成熟分生孢子中发现有少量纤维素的存在。     

DIFFERENTIATION AMONG NINE POPULATIONS OF PHLOX. I. ELECTROPHORETIC AND QUANTITATIVE VARIATION

The organization of genetic variation in Phlox drummondii was investigated using both allozyme electrophoresis and quantitative genetics. Variation at five polymorphic enzyme loci was characterized in nine populations, and variation in 16 morphological and life-history characters was examined using an analysis of full- and half-sibs in seven populations. Significant levels of genetic variation were found at enzyme loci and for metric characters. Significant heritabilities were observed for 15 of the 16 characters examined. Genetic differences among populations were revealed both by Nei's genetic distance and by phenotypic differences, summarized by discriminant analysis. Partitioning variance in allozyme frequencies among hierarchical levels of genetic organization indicated that 94% of this variance lay within populations, 4% between populations within varieties, and 2% between varieties. Partitioning phenotypic variance for metric characters indicated that 73% lay within populations, 24% lay between populations within varieties, and 3% lay between varieties. Thus, both electrophoretic and metric characters indicated that despite extensive genetic differentiation among populations, most of the evolutionary potential of the species lies within populations.     

濒危植物马蹄参枝叶形态变异与环境因子的相关性分析

调查马蹄参在湖南、广东、广西、云南4省9个天然种群20项枝叶的形态指标在种群内和种群间的表型变异,运用相关分析揭示枝和叶形态指标间以及与环境因子之间的相关性,并进行聚类分析(UPGMA)和主成分分析.结果表明:各性状在种群内和种群间均存在一定变异性.广西花坪自然保护区两个种群(HP1,HP2)枝叶形态性状(尤其是枝)的...