The Karyotype of Chi-lin Fish(Varicorhinus macrolepis) from Taishan Mountain

[Objective] The paper was to study the karyotype of Chi-lin fish （Varicorhi- nus macrolepis） from Taishan mountain. [Method] The chromosome number and karyotype of an endemic Chi-lin fish were studied using the head kidney cells. [Re- sult] The diploid chromosome number （2n） of Chi-lin fish was 50, and the karyotype of V. macrolepis was 2n=50,16m＋14sm＋20t, NF =80. No sex-chromosome was found in Chi-lin fish. [Conclusion] A detailed karyotype of this endemic cyprinid fish of Taishan Mountain was established for the first time in this study.

Isozyme Phenotypic Difference of Varicorhinus macrolepis Malic Enzyme

By using polyacrylamide gel electrophoresis and biochemical straining method, malic enzyme isozyme in 10 organs of male or female Varicorhinus macrolepis were analyzed. The results showed that malic enzyme exists in heart, gonads, eyes and gills tissues in both female and male V. macrolepis. There was only one ME-2 in heart and ME-1 in eye of female fish, while there were ME-1 and ME-2 in heart and ME-2 in eyes of male fish. Ovary contained ME-1 and ME-2, while spermary only contained ME-1. In kidney and spleen tissues, malic enzyme was found only in female fish, while in muscle, malic enzyme existed only in male fish. The phenotypic of malic enzyme had obvious differences no matter in the same organ of different sexes in V. macrolepis or different organs in the same sex. But there were no differences among different individuals of the same sex. The results will provide basic data for developmental genetics, breed improvement and directed breeding of V. macrolepis, and lay a foundation for its development and protection.

Research on Sex and Tissue Differences in Isozymic Phenotype of Varicorhinus macrolepis through Isozyme Electrophoresis

Polyacrylamide gel electrophoresis (PAGE) and biochemical staining method were used in this study for the analysis on malate dehydrogenase (MDH,E.C. 1.1.1.37) isozymes zymogram in 11 different types of tissues of male and female Varicorhinus macrolepis. It had been found for the first time that the phenotype of malate dehydrogenase (MDH),acid phosphatase (ACP) and superoxide dismutase (SOD) showed difference between male and female V. macrolepis,and there was no difference among different individuals in the same sex. Therefore,the electrophoresis band of malate dehydrogenase,acid phosphatase and superoxide dismutase could be used as an indicator for the identification of gender and tissues of V. macrolepis,which would provide basic data for the developmental genetics,variety improvement and directed breeding of V. macrolepis groups,thus facilitating the development and protection of this valuable fish species.

Research on the Lectin in Muscle Tissue of Varicorhinus macrolepis

[Objective] The paper aimed at researching lectins in muscles of Varicorhinus macrolepis and providing scientific basis for researching the adaptation mechanism and immune response of V.macrolepis to environment,which were advantageous for the protection and reproduction of V.macrolepis.[Method] V.macrolepis was used as test materials for the hemagglutination test by dialdehyde fixation to prove the existence of lectins in muscle crude homogenate of Varicorhinus macrolepis and study the physical and chemical characters.[Result] Lectins in muscle crude homogenate of V.macrolepis had shown hemagglutination effects on erythrocytes of six types of animals and had the maximum hemagglutination activity against rabbit erythrocytes,which belonged to the S-type lectins with optimal pH ranged from 4 to 8 and optimal temperature at 60 ℃.Results from the saccharide inhibition test had indicated that the sucrose was the only kind of saccharide which had inhibited the hemagglutination,suggesting that sucrose had played an important role in the process of recognition and aggregation of lectins.[Conclusion] It had been speculated that the optimal pH ranges for thermal sensitivity and hemagglutination activity of lectins in different types of aquatic organisms were similar.

泰山螭霖鱼(Varicorhinus macrolepis)在年周期中卵巢发育的组织学研究

本文对泰山螭霖鱼(Varicorhinus macrolepis)卵巢的组织结构在年周期中的变化规律进行了研究。结果表明: 泰山螭霖鱼卵母细胞的发育过程在年周期中分为6个时相。卵巢发育分期依卵巢切面中所占面积超过50%或居最高比例的卵母细胞的时相来划分,分期情况如下:2-4月份,卵巢由Ⅳ期发育至第Ⅴ期,4月份进入繁殖期;4-6 月份,为第Ⅴ期;6月下旬-7月份为第Ⅵ期,是排卵后退化阶段;8月份,为Ⅱ期卵巢(休整期);9月份-翌年的1 月份为第Ⅲ期,并以Ⅲ期卵巢越冬。由研究结果可知:泰山螭霖鱼为短期分批产卵类型,一年产卵一次。

Research on Activity and Expression of Tyrosinase in Varicorhinus macrolepis

[Objective] This study aimed to analyze tyrosinase activity and its expression in Varicorhinus macrolepis. [Method] V. macrolepis was used as experimental material for the analysis and research of tyrosinase in nine kinds of organs and tissues of male and female V. macrolepis individuals by using polyacrylamide gel electrophoresis and biochemical staining method, spectrophotometry and enzyme histochemical technology. [Result] Tyrosinase exists in the liver and pancreas, intestine and spleen of female and male V. macrolepis and in the gallbladder of male V. macrolepis. Tyrosinase activities in various tissues of V. macrolepis varied largely. Specifically, tyrosinase activities in the spleen was the maximum, which was higher in female V. macrolepis than in males. According to the enzyme histochemistry results, strong positive signals of tyrosinase existed in the spleen, intestine, liver and pancreas and gallbladder of V. macrolepis, which was the strongest in the spleen. [Conclusion] In this paper, research on tissue localization of tyrosinase in V. macrolepis had been first reported, which provided theoretical basis for further exploring the functions of tyrosinase in V. macrolepis.

Protection and Development of Varicorhinus macrolepis Germplasm Resources

Varicorhinus macrolepis is a unique rare fish in the Mount Tai area and belongs to the national second-class protected animals. Due to the development of Mount Tai area and the impact of human activities, the amount of wild V. macrolepis resources has been reduced sharply and is about to go extinct. In order to protect this rare freshwater fish, various protection policies have been promulgat- ed by various levels of government and various protection measures have been im- plemented, a number of studies have been conducted by all levels of scientific re- search institutes, and local enterprises, farmers and social groups are also actively involved in the protection activities. The protective measures and the achieved re- sults were summed up in this paper. It is believed that with the active participation of government at all levels, scientific research institutes at all levels, breeding units and social groups, significant results have been obtained after years of implementa- tion of the protection-development pattern, the amount of V. macrolepis germplasm resources has been recovered significantly, and V. macrolepis has been utilized suc- cessfully in the nutrition, health care, tourism, culture and other areas. The protec- tion model for V. macrolepis has a certain reference value for germplasm resources protection, propagation and development of other rare species.

Effects of dietary lipid levels on lipid accumulation and health status of adult Onychostoma macrolepis

The effects of different dietary lipid levels on lipid accumulation,inflammatory response,serum bio-chemical index and histological features of intestine and hepatopancreas of O.macrolepis was experimentally evaluated in an eight-weeks study.Fish(initial weight 50.11±2.86 g)were fed with five iso-nitrogenous diets(around 390 g/kg protein)varying with lipid level(5%,7%,9%,11%,13%,being LL5,LL7,LL9,LL11,and LL13 respectively)in triplicates.Results showed that the content of crude lipid in carcass and hepatopancreas were not affected by dietary lipid levels(P>0.05).Serum ALT,TP,HDL-c and MDA,etc.,were not significantly affected by diets(P>0.05),while serum total antioxidant capacity in LL9 and LL11 groups were significantly higher than the other groups(P<0.05).Histological features of hepatopancreas and intestine showed no significant difference among the five diets(P>0.05),while the height of intestine villus showed the higher trend in LL9 compared with other groups.The relative expression of lipid metabolism related genes(pparα,cpt-1α,fas,and hsl)and immune response related genes(tlr 22,nrf 2,tnfα,and il-γ)in the hepatopancreas of fish fed diets differing in their crude lipid levels were not significantly different(P>0.05).The results suggested that a proper dietary lipid level of 9%-11%could maintain higher antioxidant and health status of adult O.macrolepis.

日粮中碳水化合物类型对多鳞白甲鱼幼鱼生长性能、体组成及健康状况的影响

为探究多鳞白甲鱼幼鱼日粮中最佳碳水化合物添加类型,以葡萄糖、麦芽糖、糊精、玉米淀粉及纤维素5种碳水化合物为糖源,分别配制5种日粮,对多鳞白甲鱼幼鱼进行了为期56 d的养殖实验。结果显示,①生长及饲料利用率方面,麦芽糖组的终末体重、特定生长率、饵料系数以及蛋白质效率等指标最佳,纤维素组最差。②体成分方面,纤维素组全鱼粗脂肪水平最低,粗灰分水平显著高于麦芽糖组,麦芽糖组及纤维素组肌糖原含量最低,糊精组肝糖原含量最低。③主成分分析表明,C16:0、SFA、MUFA和PUFA比例对肝胰脏脂肪酸比例产生主要影响,其中,C16:0和SFA比例在麦芽糖组显著提高,MUFA比例在糊精组显著提高,而PUFA在纤维素组显著提高;C18:1n-9和MUFA比例对肌肉和脂肪组织总体脂肪酸组成产生主要影响,二者比例在麦芽糖组显著提高。④血清生化指标方面,麦芽糖组及纤维素组谷丙转氨酶活性、麦芽糖组谷草转氨酶活性显著较高,葡萄糖组及纤维素组碱性磷酸酶活性、球蛋白含量较高,而血糖含量反之。⑤抗氧化能力方面,麦芽糖组肝胰脏过氧化氢酶、总超氧化物歧化酶、谷胱甘肽过氧化物酶活性最高,纤维素组丙二醛含量最高。研究表明,麦芽糖是多鳞白甲鱼幼鱼适宜的碳水化合物源,玉米淀粉及糊精次之,纤维素及葡萄糖不是多鳞白甲鱼幼鱼适宜的碳水化合物源,实际生产中推荐玉米淀粉作为碳水化合物源。本研究将为多鳞白甲鱼专用饲料研发提供重要参考。

多鳞白甲鱼X型凝集素(OmITLN)基因克隆及在抗菌免疫中的功能研究

为探究秦巴山溪半洞穴鱼类——多鳞白甲鱼的先天性免疫因子X型凝集素的免疫功能,研究从多鳞白甲鱼(Onychostoma macrolepis)肝脏转录组数据库筛选出一种Intelectin基因片段,通过克隆鉴定获得Intelectin基因cDNA序列全长,命名为OmITLN;采用qRT-PCR分析OmITLN基因在健康多鳞白甲鱼8个组织和嗜水气单胞菌感染后两个免疫组织的表达情况;构建OmITLN基因的原核表达载体并在大肠杆菌BL21中诱导表达,获得OmITLN重组蛋白,采用ELISA法以及荧光标记法检测OmITLN对病原菌的凝集效果及糖结合情况。结果显示,OmITLN是一种X型凝集素基因,其cDNA全长960 bp,编码315个氨基酸。OmITLN蛋白含有1个N端的纤维蛋白原相关结构域(Fibrinogen-Related Domain,FReD)和1个C端Intelectin特异性区域,不具有信号肽和跨膜区。系统进化树分析发现,OmITLN与鲫、团头鲂、鳙、草鱼等鲤科鱼类的Intelectin聚为一支,亲缘关系最近。OmITLN基因在所检测组织均有表达,且在脾脏和肝脏中的表达量最高。腹腔注射1.65×107 CFU/尾的嗜水气单胞菌后,OmITLN基因在感染鱼的脾脏和肝脏组织的表达量均呈先上调后下降的趋势,但表达量达到最高值的时间不同,表明其两个组织对嗜水气单胞菌感染均有不同程度的免疫响应。试验利用原核表达系统成功纯化获得体外重组蛋白OmITLN,OmITLN重组蛋白对副溶血弧菌、嗜水气单胞菌、大肠杆菌、无乳链球菌、蜡状芽孢杆菌和金黄色葡萄球菌均具有不同程度凝集作用。OmITLN重组蛋白对D-乳糖的结合活性最好,木糖、D-半乳糖、蔗糖和D-葡萄糖的结合活性次之,对海藻糖、D-果糖和D-麦芽糖没有结合活性;对肽聚糖(PGN)的结合活性高于脂多糖(LPS)。OmITLN作为重要的模式识别受体,参与了多鳞白甲鱼应答病原菌感染的防御过程。

贵州省台江县南宫自然保护区翠柏群落特征与多样性研究

为了解翠柏群落结构特征和多样性状况,更好地保护和利用翠柏资源,采用样地调查的方法对贵州省台江县南宫自然保护区翠柏群落进行调查。结果表明:(1)群落有植物41科51属64种(含种以下等级),其中蕨类植物5科5属6种;种子植物36科46属58种;(2)群落的翠柏(0.4335)重要值最大,次之为南方红豆杉(0.1041),翠柏是建群种,南方红豆杉、枫香、杉木为乔木层优势种;群落更新能力差,存在年龄断层现象,是个衰退型群落;(3)群落丰富度指数灌木层>草本层>乔木层;优势度指数乔木层>草本层>灌木层;H多样性指数草本层>乔木层>灌木层;均匀度指数乔木层>草本层>灌木层。说明灌木层物种丰富度最明显;乔木层优势度最明显;草本层H多样性指数最高,乔木层次之;乔木层均匀度指数最大,草本层次之;(4)本群落与广西木论、云南易门等地的翠柏群落具有相似性。南宫翠柏群落是一个古老的群落,应加大对其的保护。

秦巴山区多鳞铲颌鱼种质资源调查及保护对策研究

秦巴山区多鳞铲颌鱼主要分布在汉水水系的中上游高海拔水域,对秦巴山区多鳞铲颌鱼(Varicorhinus macrolepis)的生境、分布区、生物学特性、繁殖特性进行了调查。结果表明,安康地区各主要河流建有大量水利工程及拦河坝,已建小水电站86座(含2座已废弃),在建11座,规划待建42座,建有拦河坝及水库72座。多鳞铲颌鱼属冷水性、杂食性鱼类,性别比接近1∶1,分批产卵。紫阳、平利、镇坪、宁陕等县多鳞铲颌鱼分布较多。紫阳县任河干流及其支流中的麻柳河、盘厢河、朱溪河、绕溪河等水域内该种群属优势种群,镇坪县南江河、岚河支流正阳河也有分布,而白河县白石河、冷水河,旬阳县,汉滨区分布较少。人类破坏、生境破碎化、生境退化和污染、过度利用是多鳞铲颌鱼濒危的主要原因,可采取原地保护、迁地保护、宣传教育、加强渔政执法、加强研究等措施来保护和恢复种群数量,达到永续利用的目的。

泰山赤鳞鱼的胚胎发育和仔鱼发育

试验结果表明:泰山赤鳞鱼产卵时间在4～5月,自然产卵水温为18～26℃,产卵于水底细沙中,分散产卵.卵呈弱粘性,金黄色,属沉性卵,平均卵径2.5 mm.在水温18～22℃条件下,胚胎从受精到孵化出膜历时66 h,经过了胚盘形成期、卵裂期、囊胚期、原肠胚期、神经胚期、器官分化期6个主要阶段.初孵仔鱼全长6.0～6.5 mm.卵黄囊前部膨大成球形,约占鱼体整个长度的1/3;后端为均匀的棒状.仔鱼孵出到消化道完全形成约经历136 h.

濒危植物翠柏的个体生长动态及种群结构与种内竞争

在样地调查的基础上,对滇中高原地区翠柏个体生长状况以及不同郁闭度翠柏纯林的种群结构、种内竞争进行比较研究。结果表明:1)翠柏植株胸高断面积与树高之间存在着显著的指数函数关系,胸径和树高分别为31～40cm和19m左右时是其生长过程中较为明显的转折点,之前,树高生长占有明显的优势,随后,胸径增长迅速,树高生长速度减慢,直至停止;2)滇中地区翠柏天然林林地破碎化程度较高,径级结构的分布极不规律,呈现出明显的不连续状;3)现有零星分布的翠柏纯林中,植株密度和大小存在较大差异,以致种群中单株平均竞争强度压力差异较大;同时,林下幼小植株受到的种内竞争压力极为强烈,但随着胸径的增大,植株受到的竞争压力逐渐减小,并在胸径31～40cm时形成明显拐点。胸径小于31～40cm的植株受到的竞争压力较大,且随胸径的增大出现急剧下降;胸径大的植株,如大于37cm,受到的竞争压力较小,且随胸径的增大,竞争强度的变化较为平缓。在翠柏的保育管理过程中,加强对竞争压力较小的林缘、林隙等地区的管理有助于拓展翠柏生存范围。同时,在经营和利用翠柏植株资源时,可以参照其天然植株生长特征,对林分进行合理的经营和管理。

多鳞铲颌鱼性腺分化发育的组织学研究

用光镜和透射电镜对多鳞铲颌鱼(Varicorhinus macrolepis)性腺的分化发育进行组织学研究,观察了雌雄性腺的形成过程及各级生殖细胞的形态特征。结果表明,多鳞铲颌鱼卵巢和精巢开始分化的时间分别在80日龄和90日龄,雌鱼和雄鱼的性成熟年龄分别为36月龄和24月龄,精子和卵子的发育均可分为5个时期。在多鳞铲颌鱼的生长发育过程中,12月龄(雄鱼)和30月龄(雌鱼)性腺质量与体长、体质量之间具有极显著的相关关系。

木论喀斯特林区翠柏群落学特征的初步研究

广西木论喀斯特林区，位于我国中亚热带南缘。据６００ｍ２翠柏群落样方调查结果：有维管束植物７６种，隶５１科６９属，其中单种的属占８９．９％。植物区系以热带分布区类型为主，占总属数（不含世界分布）的７１．０％，温带分布占２５．８％，但群落的种类组成是以亚热带成分占优势，按Ｒａｒｎｋｉａｅｒ生活型分类系统，高位芽植物占８２．９％，地面芽植物占１５．８％，一年生植物占１．３％，缺少地上芽和地下芽植物；从生理角度分析，叶级以小型叶（包括微型和鳞型叶）为主，占５９．２％；叶型以单叶占的比例最大，为８６．８％，群落结构可分为乔、灌、草三层，其中乔木层又可分出二个亚层。对该群落各乔木种群的大小结构分析，显示出翠柏群落是相对稳定性较大的群落。

广西翠柏属(柏科)植物小志

订正了广西翠柏属(柏科)植物,确定了广西分布有2种,即翠柏和岩生翠柏,其中分布于广西石灰岩山顶的翠柏属植物为岩生翠柏,该种与翠柏不同在于鳞叶先端钝状或宽钝状,球果宽卵形,具极短的柄,种鳞4,可育种鳞先端弯曲、圆形,表面粗糙,无尖头。最后根据IUCN的评估标准对广西翠柏属植物进行了濒危等级的评估,并提供了两种植物的地理分布信息和野外生态照片。

濒危植物翠柏个体生长特性研究

濒危植物翠柏为柏科高大珍贵用材树种,通过对翠柏主要分布区墨江、昌宁翠柏植株树干解析,研究了两地翠柏的生长特性,结果表明:(1)翠柏在20年前生长缓慢,20—60年进入快速生长期,70年以后达到数量成熟,材积生长量开始下降并趋于稳定;(2)翠柏幼苗期具有耐荫性和长大后有喜光特性,墨江地区翠柏生长好于昌宁地区,其生长表现与两地自然条件相符合;(3)针对翠柏特点,提出幼苗时搭建荫棚,幼树时期进行疏伐,数量成熟时期进行间伐或疏伐等促进生长经营措施,为翠柏资源培育与保护奠定基础。

泰山赤磷鱼酯酶同工酶(EST)的研究

运用垂直板聚丙烯酰胺凝胶电泳技术 ,对泰山赤磷鱼早期发育不同时期(未受精卵、受精卵、原肠晚期、色素期 )和成体眼、脑、心、肌、肝、肾 6种组织酯酶进行研究 ,结果表明 :(1 )泰山赤磷鱼酯酶同工酶在未受精卵和早期胚胎发育时期均表达 ,并且酶谱和酶活性有差异 ,该酶的激活时序与动物胚胎发育中的细胞分化密切相关 ;(2 )泰山赤磷鱼酯酶同工酶具有明显的组织特异性 ,酯酶同工酶基因表达与组织来源和细胞分化密切相关 .