密斑刺鲀(Diodon hystrix)fshβ和lhβ基因的克隆及表达分析

密斑刺鲀(Diodon hystrix)是南海名特优鱼类,同时具备食用和中药保健价值,但目前关于密斑刺鲀生殖生理的研究较少,影响了该鱼繁育理论研究的发展。众所周知,促性腺激素(Gonadotropic hormone,GtH)是生殖调控的关键激素之一,在生殖调控轴中发挥着承上启下的作用。利用转录组分析及分子克隆技术从密斑刺鲀中克隆并鉴定出fshβ和lhβ基因的cDNA序列,通过生物信息学分析,发现密斑刺鲀fshβcDNA开放阅读框共363 bp,编码120个氨基酸;lhβcDNA开放阅读框共420 bp,编码139个氨基酸。通过序列比对和系统进化树分析,表明密斑刺鲀Fshβ和Lhβ均与鲀科类的鱼的亲缘关系较近。利用三级结构分析,发现Fshβ的三级结构相对保守,而Lhβ的三级结构差异较大。利用实时荧光定量的方法检测了密斑刺鲀fshβ和lhβ基因的组织分布情况,结果表明密斑刺鲀fshβ和lhβ在垂体的表达量最高,其次是性腺。此外,进一步检测了fshβ和lhβ基因在雌雄性腺发育过程中的表达模式,发现垂体中fshβ基因伴随性腺的发育,表达量逐渐下降,而lhβ基因的表达量呈逐渐增加模式。研究结果将为构建密斑刺鲀生殖调控网络提供研究基础。

Hystrix patula与Pseudoroegneria libanotica属间杂种的细胞学研究

为研究猬草Hystrixpatula的染色体组组成,进行了H.patula与Pseudoroegnerialibanotica的人工杂交,获得杂种F1,观察了亲本和杂种F1花粉母细胞减数分裂染色体配对行为。杂种F1染色体配对较高,84%的细胞形成7个或7个以上二价体,其构型为6.08Ⅰ+7.48Ⅱ,C-值为0.69。结果表明,H.patula含有St染色体组。

Genetic Diversity of Wild and Cultivated Populations of Castanopsis hystrix in China

[Objective] This study aimed to investigate the genetic variation of wild and cultivated populations of Castanopsis hystrix. [Method] Genetic variation of five wild populations and three cultivated populations of Castanopsis hystrix, was investigated with ISSR-PCR amplification. Totally, 151 individuals were selected and analyzed by amplification using nine pairs of ISSR primers screened. [Result] Each primer pair produced 7-20 bands and 122 polymorphic bands were obtained. At population level, ISSR diversity in the wild populations (P=59.84%, HPOP=0.182 7, I=0.285 6) was higher than which in cultivated ones (P=54.87%, HPOP=0.136 6, and I=0.219 8). The genetic differentiation coefficient among wild populations (GST) was 0.99. The similar population structure was found in three cultivated populations (GST=0.127 5). According to the UPGMA cluster analysis, the genetic distance among wild populations became larger with the increase of geographical distance. [Conclusion] Compared with other seed plants, with either a similar life history or various breeding system attributes, relatively low level of genetic diversity was observed in these five wild populations, which was caused by population size reduction and habitat fragmentation related to human activities. The formation of population structure may be explained by the species’ breeding system.

密斑刺鲀(Diodon hystrix)gnrh基因的克隆及表达分析

密斑鲀刺(Diodon hystrix)不仅具有食用价值,同时也是传统的中药及保健食品,但目前关于密斑刺鲀的生殖内分泌调控研究较少,限制鲀了密斑刺的人工繁育基础理论的发展。总所周知,促性腺激素释放激素(gonadotropin-releasing hormone,GnRH)是生殖内分泌调控的关键因子之一。利用转录组分析及分子克隆技术,获得密斑刺鲀促性腺激素释放激素(gonadotropin-releasing hormone,GnRH)gnrh2和gnrh3基因的cDNA开放阅读框(Open Reading Frame,ORF)序列,并利用生物信息学的方法,获鲀得密斑刺gnrh2 cDNA开放阅读框共279 bp,可编码92个氨基酸残基;密斑鲀刺gnrh3 cDNA开放阅读框共270 bp,可编码89个氨基酸残基。通过系统进化分析表明,密斑鲀刺GnRH与鲀科类的鱼具有较近的亲缘关系。通过聚合酶链式反应(Polymerase Chain Reaction,PCR)半定量的方法检测gnrh2和gnrh3基因在密斑刺鲀各组织中的分布,结果发现gnrh2和gnrh3基因在脾、鳃和垂体中的表达量较高,而在心脏中的表达量最低。此外,利用鲀实时荧光定量的方法检测了密斑刺gnrh2和gnrh3基因在性腺发育不同时期的表达模式,结果显鲀示密斑刺gnrh2基因在不同的性腺发育时期中没有显著性的变化,而gnrh3基因随着性腺发育成熟而不断升高。研究主要获得两个创新性的重要结果:首鲀先是证实了密斑刺存在两种gnrh基因(gnrh2和gnrh3),其次是提示了gnrh3基因可能是鲀密斑刺生殖调控的主要gnrh类型。研究结果可为深鲀入研究密斑刺生殖内分泌调控机理提供研究方向和基础。

Preliminary Study on the Adaptability of Cucumis hystrix Chakr. under Aluminum Salt Stress

[Objective] The study was carried out to research the adaptability of Cucumis hystrix Chakr.under aluminum salt stress.[Method] Using Cucumis hystrix Chakr.and three cultivated cucumbers as tested materials,the change of physiological indexes including POD activity,SOD activity,electrolyte leakage and the content of malondialdehyde（MDA）,proline（Pro）content and soluble sugar were studied.[Result] The POD activity,SOD activity,proline content and soluble sugar content of Cucumis hystrix were higher than these of cultivated cucumbers,while electrolyte leakage and MDA content were lower,and it showed that Cucumis hystrix had better adaptability under aluminum salt stress.[Conclusion] Our study could lay a foundation for the improvement of cultivated cucumber by using the excellent characteristics of Cucumis hystrix.

Molecular Identification of Castanopsis hystrix,Castanopsis carlesii and Quercus griffithii Using ISSR-PCR Method

[Objective] This research aimed to develop molecular identification method for Castanopsis hystrix,Castanopsis carlesii and Quercus griffithii.[Method] DNA fingerprints of C.hystrix,C.carlesii and Q.griffithii were established by using ISSR-PCR method.Cluster Analysis was carried out by using UPGMA method based on Nei＇s genetic distances among each individual.[Result] Six polymorphic primers were selected from 50 ISSR primers for ISSR-PCR amplification,and totally 86 discernible DNA bands were amplified with 53 polymorphic bands,accounting for 61.2% of the total.The average number of DNA bands amplified by each primer was 10.75.Specifically,totally 5 primers had amplified differential bands and specific bands,which were able to accurately identify C.hystrix,C.carlesii and Q.griffithii.As calculated by DPS v3.01 software,the genetic distances among test materials were ranged from 0.166 67 to 0.809 52,with an average of 0.563 57.[Conclusion] ISSR-PCR method can be used to identify C.hystrix,C.carlesii and Q.griffithii effectively.

小麦族下Hystrix longearistata和Hystrix duthiei的生物系统学研究

对Ｈｙｓｔｒｉｘｄｕｔｈｉｅｉ、Ｈ．ｌｏｎｇｅａｒｉｓｔａｔａ和它们的人工种间杂种花粉母细胞减数分裂染色体配对行为、繁育特征和形态特征进行了比较分析，结果表明：（１）这两个分类单位形态差异较小，Ｈ．ｌｏｎｇｅａｒｉｓｔａｔａ的外稃芒较长，叶片较宽，每小穗具２～３个小花；Ｈ．ｄｕｔｈｉｅｉ的外稃芒较短，叶片较窄，每小穗具１～２个小花。（２）它们很容易杂交，杂种Ｆ１染色体配对频率很高，为１３～１４个二价体。（３）亲本种花粉育性和结实性正常，杂种Ｆ１花粉育性较低，结实性较差。（４）Ｈ．ｌｏｎｇｅａｒｉｓｔａｔａ和Ｈ．ｄｕｔｈｉｅｉ亲缘关系很近，是同一物种。由于地理分布和生境的差异，使它们在形态上开始分异，并出现一定程度的生殖隔离。把Ｈ．ｌｏｎｇｅａｒｉｓｔａｔａ处理为Ｈｙｓｔｒｉｘｄｕｔｈｉｅｉ的一亚种是合理的。

野黄瓜(Cucum is hystrix Chakr.)与3种不同基因型栽培黄瓜(C. sativus L.)种间杂交及杂种鉴定

野黄瓜(Cucum is hystrix,2n=24)为短日照植物,因其与栽培黄瓜(C.sativus,2n=14)花期不遇等杂交障碍的存在而使种间杂交难以顺利进行。对野黄瓜短光照处理40 d以上,使得两者花期相遇,成功地进行了野黄瓜与3种不同基因型栽培黄瓜(“二早子”、“长春密刺”和“NC4406”)的正反杂交,从中获得了3种杂交组合的幼胚。采用改良的方法对这3种基因型杂种胚进行胚胎拯救,再生频率达80%以上,其杂种的真实性通过植物学性状观察、体细胞染色体计数和天冬氨酸转氨酶(AAT)同工酶分析得到了证实。研究中还发现不同基因型杂种F1的育性存在明显差异,其中以C.hystrixדNC4406”育性最高,花粉可染率达23.3%,是在二倍体水平上进行育种利用的良好材料。

刺栲(Castanopsis hystrix)的气体交换和叶绿素a荧光特征

用LI-6400便携式光合测定系统和PAM-2100便携式植物荧光分析仪分别在初夏和秋季测定了广州龙眼洞林场刺栲(Castanopsishystrix)幼树叶片的气体交换和叶绿素a荧光参数。结果表明:1)刺栲冠层阳生叶的光饱和点为400μmolphotonsm-2s-1,最大光合速率在4-6μmolm-2s-1之间,说明刺栲幼树保持阳生植物的特征。阴生叶与阳生叶的叶绿素a/b值相似,均约为2.4,而阴生叶叶绿素b含量高于阳生叶,表明阴生叶具有潜在利用弱光的能力;2)刺栲幼树冠层叶片的PSⅡ的最大光化学效率Fv/Fm和光化学量子产额(Yield)的日动态变化在中午略有下降,说明其在强光下能发生可逆性的光抑制。刺栲冠层阳生叶的最大光合能力比较低,整个冠层叶能高效而稳定地利用散射和透射光,这些特征使得常绿的刺栲幼树能在林边或林下良好生长,较好地适应不同的光环境并成功渡过幼苗期,最终成为群落中的上层优势种。

周口店田园洞古人类化石点的无颈鬃豪猪(Hystrix subcristata)

周口店田园洞位于北京猿人遗址西南6km处,是新发现的晚期智人化石地点。该地点是迄今我国北方地区第四纪化石地点中含豪猪材料最丰富的。从头骨、牙齿、头后骨骼的形态特征及大小判断,田园洞的豪猪应归入我国现生无颈鬃豪猪(Hystrixsubcristata)。14C测年表明,含豪猪化石的地层时代为3.05～0.467万年,为更新世末期到全新世中期,是目前所知豪猪化石在我国北方地区的最晚记录。

Asynchronous meiosis in Cucumis hystrix–cucumber synthetic tetraploids resulting in low male fertility

Interspecific hybridization and allopolyploidization contribute to the improvement of many important crops. Recently, we successfully developed an amphidiploid from an interspecific cross between cucumber(Cucumis sativus, 2n = 2x = 14) and its relative C. hystrix(2n = 2x = 24) followed by chemical induction of chromosome doubling. The resulting allotetraploid plant was self-pollinated for three generations. The fertility and seed set of the amphidiploid plants were very low. In this study, we investigated the meiotic chromosome behavior in pollen mother cells with the aid of fluorescence in situ hybridization, aiming to identify the reasons for the low fertility and seed set in the amphidiploid plants. Homologous chromosome pairing appeared normal, but chromosome laggards were common, owing primarily to asynchronous meiosis of chromosomes from the two donor genomes. We suggest that asynchronous meiotic rhythm between the two parental genomes is the main reason for the low fertility and low seed set of the C. hystrix–cucumber amphidiploid plants.

Hepatoprotective effect of leaf extracts from Citrus hystrix and C.maxima against paracetamol induced liver injury in rats

The present investigation is aimed to evaluate the hepatoprotective effects of Citrus hystrix and Citrus maxima(Red and White variety)methanolic leaf extracts on paracetamol induced toxicity.Leaf extracts were given in the dose of 200 mg/kg body weight for 7 days and toxicity was induced by paracetamol(2 g/kg)on day 5.Silymarin(100 mg/kg body weight)was used as reference standard.On the 7th day animals were sacrificed and liver function markers(ALT,AST,ALP),total bilirubin and total protein in blood serums and hepatic antioxidants(SOD,CAT,GSH and GPx)in liver homogenate were estimated.The leaf extracts restored the liver function markers and hepatic antioxidants to the normal level than elevated levels noticed on paracetamol control at P<0.001.Reversal of hepatoarchitecture has also been registered.The present study shows that C.hystrix and C.maxima leaf extracts possess hepatoprotective action against paracetamol induced hepatotoxicity.©2015 Beijing Academy of Food Sciences.Production and hosting by Elsevier B.V.All rights reserved.

Cardioprotective and hepatoprotective effects of Citrus hystrix peels extract on rats model

Objective:To observe the combination effect of doxorubicin and Citrus hystrix(kaffir lime's)peel ethanolic extract(ChEE)on blood serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST)activity and cardio-hepato-histopathology of female Sprague Dawley rats.Methods:Doxorubicin and ChEE(5 rats per group)were administered in five groups of 3 rats each for 11 d.Group I:doxorubicin(dox)4.67 mg/kg body weight;Group II:dox+ChEE 500 mg/kg body weight;Group HI:dox+ChEE 1000 mg/kg body weight;Group IV:ChEE 1000 mg/kg body weight;Group V:untreated(control).Results:ChEE repaired cardiohistopathology profile of doxorubicin induced cardiotoxicity and hepatotoxicity rats,but did not repair neither hepatohistopathology profile nor reduce serum activity of ALT and AST.Conclusion:ChEE has potency to be developed as cardioprotector agent in chemotherapy.

基于Hystrix服务熔断降级策略的优化分析与研究

本方案公开了一种基于Hystrix框架的动态熔断判断方法,通过压力测试得出当前服务业务的最大负载量max F,通过测试得出不同压力下的业务失败率范围,改造Hystrix框架源码,重写Health Count类,记录请求运行总数与运行错误总数,重写PLUS统计方法,记录数据,改造Hystrix框架源码,重写断路器实现类,根据上一步统计的相关数据计算出服务负载量占最大负载量的百分比,来动态得出熔断失败率算法,最终根据得出的动态失败率来决定是否需要进行熔断。本方案主要对源码中的请求量统计进行重写,记录请求的相关统计数,同时对断路器实现部分重写断路状态判断调整算法,以实现动态熔断规则的处理。

Relationship between the stoichiometric characteristics and leaf functional traits of Castanopsis hystrix in different ages

Both the stoichiometric characteristics and leaf functional traits can reflect the adaptability of plants to changes in the external living environment.In particular,for varying aged forest plants,the study of stoichiometric characteristics and leaf functional traits can reflect the plant’s life history strategy and its resource investment and allocation methods.In this paper,the 1.5-year-old,5-year-old,15-year-old and 20-year-old Castanopsis hystrix were selected as the research objects to investigate the trends of stoichiometry and leaf functional traits,and their synergistic changes were verified.The results showed that with the increase of age,the stability of C.hystrix stoichiometry gradually increased,and the functional traits also tended to invest in security.However,there was no synergistic change between the two,which might be attributed to the different sensitivity of the C.hystrix leaf to the environment at the life history level of 1.5-year-old to 20-year-old.

Cytological Analyses on Development of Male and Female Gametophytes in an Interspecific Hybrid F_1 from Cucumis hystrix Chakr. × Cucumis sativus L.

An interspecific hybrid F1 of Cucumis hystrix Chakr. x Cucumis sativus L. （NC4406） was used to establish the developmental sequence and to characterize the male and female gametophytes at cytological level for further understanding of the phylogenic relationship and the mechanism of fertility or sterility in the interspecific hybrid F1 The development of male and female gametophytes was studied through meiotic analysis and paraffin section observation technique, respectively. Meanwhile, the fertility level was assessed through hybrid F1 backcrossing to cultivated cucumber 4406. Variable chromosome configurations were observed in the pollen mother cells （PMCs） of hybrid F1 at metaphase Ⅰ , e.g., univalents, bivalents, trivalents, quadravalents, etc. At anaphase Ⅰ and Ⅱ, chromosome lagging and bridges were frequently observed as well, which led to the formation of polyads and only a partial number of microspores could develop into fertile pollen grains （about 23.3%）. Observations of the paraffin sections showed numerous degenerated and abnormal embryo sacs during the development of female gametophytes, and only 40% of the female gametophytes could develop into normal eight-nuclear megaspore. On an average, 22.8 and 6.3 seeds per fruit could be obtained from the reciprocal backcross. The interspecific hybrid F1 of C. hystrix x NC4406 was partially fertile; however, the meiotic behaviors of hybrid F1 showed a high level of intergenomic recombination between C. hystrix and C. sativus chromosomes, which indicated that it plays an important role for introgression of useful traits from C. hystrix into C. sativus.

Ecotoxicological Effects of Cosmetic Formulas Containing Chemical and Mineral UV Filters on Seriatopora hystrix Fragments

Background: Over the last few years sunscreen products have been suspected to be harmful to corals, especially because of their putative negative impact on symbiotic microalgae housed by these cnidarians. Previous publications reported that minerals or chemical UV filters could induce the release of microalgae from corals inducing their bleaching. The study of the ecotoxicity of finished cosmetic products containing these filters is important. Objectives: We sought to assess ex vivo the toxicity of five emulsions containing UV-filters on coral cuttings of Seriatopora hystrix. Materials and Methods: Coral cuttings were put in contact with 5 different emulsions containing UV-filters. The toxicity readout was the ability to induce polyp retraction and/or fragment bleaching of the coral cuttings of Seriatopora hystrix. Results: In our experimental conditions, none of the five tested formulas neither induced any significant polyp retraction nor triggered fragment bleaching of the coral. Conclusions: The five tested emulsions containing UV-filters did not modify coral cuttings. In vivo, larger tests are necessary to verify the results of this ex vivo pilot study.

甜瓜属野生种Cucumis hystrix Chakr.胞质葡萄糖-6-磷酸脱氢酶基因的克隆与启动子结构分析

根据植物葡萄糖-6-磷酸脱氢酶(G6PDH,EC1.1.1.49)氨基酸保守区域,设计简并引物,采用RT-PCR技术克隆得到甜瓜属野生种Cucumis hystrix Chakr.(2n=24)G6PDH基因cDNA片段;随后基于该序列设计特异引物,PCR方法筛选野生种BAC文库,获得2个BAC阳性单克隆。测序后获得了G6PDH基因全序列及其上游启动子序列,GenBank登录号:JQ771576。序列分析显示:G6PDH基因全长约6.5 kb,由15个外显子和14个内含子组成;内含子序列均符合5'-gt-ag-3'结构。外显子拼接后获得了G6PDH基因的开放阅读框(ORF)序列,序列全长1 551 bp,编码516个氨基酸,与黄瓜基因组网站公布的G6PDH基因核苷酸序列和氨基酸序列同源性分别为98.71%和99.03%。野生种G6PDH氨基酸序列N端缺少转运肽序列,确定为野生种胞质G6PDH。氨基酸序列与烟草、马铃薯、荷兰芹、猕猴桃、拟南芥、大豆、小麦、玉米、葡萄、杨树等植物胞质G6PDH同源性高达77%以上。系统发育树分析发现,甜瓜属胞质G6PDH与茄科植物最先聚类,植物胞质G6PDH在分子进化水平上与物种进化相符。启动子结构分析表明:序列含有启动子基本元件TATA-box、CAAT-box,还含有丰富的光响应元件,与环境胁迫响应相关的不同顺式作用元件,促进高水平转录的5UTR Py-rich stretch元件和提高表达的CAT-box元件等。

Analysis and Evaluation of Nutrient Composition in Different Tissues of the Wild Spot-fin Porcupine Fish(Diodon hystrix)

Nutrient compositions including proximate, fatty acids, amino acids, minerals and vitamins in tissues of the wild spot-fin porcupinefish(Diodon hystrix) were analyzed in detail to assess their nutrient value and to further explore the dietary requirements of this species farmed. Diodon hystrix individuals with similar body weight(284.6-358.5 g) and body length(15.9-21.8 cm) were collected from the South China Sea from Aug., 2015 to Aug., 2016. In general, skin and muscle possessed negligible fat contents(0.3-0.4 g/100 g wet weight) and low energy value(317.4-497.3 k J/100 g), while proteins(17.8-25.4 g/100 g wet weight) were found to be the richest component. The dominant amino acids in both muscle and skin of D. hystrix included histidine, cysteine, tyrosine, isoleucine, leucine and lysine, while the major mineral elements were calcium, magnesium, potassium,sodium, and phosphorus. It was apparent that D. hystrix had high content of amino acids, and the rate of essentia amino acids was accorded with FAO/WHO standard. This study illustrated that the wild D. hystrix would be a valuable natural source of protein, with high commercial potential, and especially, skin and muscle were more valuable. Moreover, the basic data from the present study would supply the fundamental reference for formulated diet and culture of this species.

A new Late Miocene Hystrix(Hystricidae,Rodentia)from Turkey

TheÇorakyerler fossil locality in Anatolia reveals unique faunal elements.This study introduces Hystrix kayae sp.nov.,a new Late Miocene porcupine initially classified as Hystrix sp.Yavuz et al.,2018.This finding expands our knowledge of Late Miocene Anatolian porcupines,bringing the total number of known species to three.H.kayae sp.nov.is larger than Hystrix aryanensis and exhibits greater upper cheek tooth crown height.Its cheek tooth morphology differs from Hystrix depereti,and it possesses distinctive U-shaped choanae unlike Hystrix primigenia.TheÇorakyerler locality may predate sites with H.aryanensis and H.primigenia but aligns temporally with Hystrix parvae localities.This study enhances our understanding of Late Miocene porcupine diversity in Anatolia,emphasizing the importance ofÇorakyerler in unraveling the evolutionary history of these fascinating mammals.