鹅掌楸属（Liriodendron L．）种质‘优酉’的遗传背景研究

应用光学显微镜和扫描电子显微镜,以鹅掌楸[Liriodendron chinense（Hemsl．）Sarg．]、北美鹅掌楸（L.tutipifera L．）及杂种鹅掌楸（L．chinense×L．tulipifera）为对照,对重庆酉阳鹅掌楸（Liriodendron sp．）种源‘优酉’（‘Youyou’）的外部形态特征、叶表皮微形态结构特征和花粉形态特征进行观察比较,并利用RAPD标记技术对‘优酉’的遗传关系进行了分析。结果表明,鹅掌楸属各树种在外部形态上存在差别,但在叶表皮和花粉形态特征上表现出明显的一致性,‘优酉’的各形态特征与鹅掌楸有较多相似性。RAPD标记分析结果表明,‘优酉’的遗传距离与鹅掌楸和北美鹅掌楸较远,但与鹅掌楸的遗传关系相对较近。推测‘优酉’可能是由较原始的鹅掌楸向较进化的北美鹅掌楸过渡的1个中间类型。‘优酉’生长速度快、观赏价值高,可直接用于园林绿化及林木种植。     

宫颈癌p16基因甲基化及表达的研究

为了探讨p16基因甲基化及异常表达在宫颈癌中的意义,分别采用甲基化特异性PCR（MSP）方法检测不同病理类型和临床分期的60例宫颈癌组织中p16基因启动子区域5´CpG岛甲基化状态;采用PCR方法检测p16基因外显子1（E1)和外显子2(E2)纯合缺失情况;采用免疫组化的方法分析p16蛋白的表达缺失和减弱情况。结果显示正常对照组织及癌旁p16基因无甲基化,且无E1和E2缺失和p16蛋白表达异常。60例宫颈癌标本的甲基化率为21.67%（13/60）;p16 基因缺失率为15.00%（9/60）;p16蛋白表达下降或无表达为51.67%（31/60）。可见p16基因蛋白的阳性表达率随着临床分期升高呈明显下降趋势。结果提示p16基因失活在宫颈癌中多见且与病理分级密切相关。p16 基因甲基化在宫颈癌发生中起着一定作用。Abstract： To detect hypermethylation and aberrant expression of the p16 gene in cervical carcinoma (CC), methylation-specific PCR (MSP) was used to determine the methylation status of 5´CpG islands of the p16 gene, loss or decrease of p16 expression was analyzed by immunohistochemistry (IHC), and homozygous deletion of exon 1 (E1) and/or exon 2 (E2) was determined by PCR. in 60 cases of CC at different pathological grades and clinical stages. The results showed absence of methylation and presence of normal expression of the p16 gene in the control and adjacent tissues of CC. Hypermethylation, loss or decrease of expression and deletion of the p16 gene were detected in 21.67%(13/60), 51.67%(31/60) and 15.00%(9/60) of the tumor tissues, respectively. The rate of p16 expression markedly reduced with the increase of clinical stages. Our data suggested that inactivation of the p16 gene was a frequent event and positively correlated with pathological grades in CC, and that methylation of the p16 gene was an important event in carcinogenesis of CC.     

Coexistence of mitochondrial 12S rRNA C1494T and CO1/tRNA(Ser(UCN)) G7444A mutations in two Han Chinese pedigrees with aminoglycoside-induced and non-syndromic hearing loss

Mutations in mitochondrial DNA are one of the important causes of hearing loss. We report here the clinical, genetic, and molecular characterization of two Han Chinese pedigrees with maternally transmitted aminoglycoside-induced and nonsyndromic bilateral hearing loss. Clinical evaluation revealed the wide range of severity, age-at-onset, and audiometric configuration of hearing impairment in matrilineal relatives in these families. The penetrances of hearing loss in these pedigrees were 20% and 18%, when aminoglycoside-induced deafness was included. When the effect of aminoglycosides was excluded, the penetrances of hearing loss in these seven pedigrees were 10% and 15%. Sequence analysis of the complete mitochondrial genomes in these pedigrees showed the presence of the deafness-associated 12S rRNA C1494T and CO1/tRNA(Ser(UCN)) G7444A mutations. Their distinct sets of mtDNA polymorphism belonged to Eastern Asian haplogroup C4a1, while other previously identified six Chinese mitochondrial genomes harboring the C1494T mutation belong to haplogroups D5a2, D, R, and F1, respectively. This suggested that the C1494T or G7444A mutation occurred sporadically and multiplied through evolution of the mitochondrial DNA (mtDNA). The absence of functionally significant mutations in tRNA and rRNAs or secondary LHON mutations in their mtDNA suggest that these mtDNA haplogroup-specific variants may not play an important role in the phenotypic expression of the 12S rRNA C1494T and CO1/tRNA(Ser(UCN)) G7444A mutations in those Chinese families. However, aminoglycosides and other nuclear modifier genes play a modifying role in the phenotypic manifestation of the C1494T mutation in these Chinese families.     

A Novel missense mutation of COL2A1 gene in a large family with stickler syndrome type I

Stickler syndrome type I (STL1, MIM 108300) is characterized by ocular, auditory, skeletal and orofacial manifestations. Nonsyndromic ocular STL1 (MIM 609508) characterized by predominantly ocular features is a subgroup of STL1, and it is inherited in an autosomal dominant manner. In this study, a novel variant c.T100>C (p.Cys34Arg) in COL2A1 related to a large nonsyndromic ocular STL1 family was identified through Exome sequencing (ES). Bioinformatics analysis indicated that the variant site was highly conserved and the pathogenic mechanism of this variant may involve in affected structure of chordin‐like cysteine‐rich (CR) repeats of ColIIA. Minigene assay indicated that this variant did not change alternative splicing of exon2 of COL2A1. Moreover, the nonsyndromic ocular STL1 family with 16 affected members showed phenotype variability and certain male gender trend. None of the family members had hearing loss. Our findings would expand the knowledge of the COL2A1 mutation spectrum, and phenotype variability associated with nonsyndromic ocular STL1. Search for genetic modifiers and related molecular pathways leading to the phenotype variation warrants further studies.     

梵净山保护区食蟹獴(Herpestes urva)栖息地选择与日活动节律

野生动物栖息地选择与日活动节律是其长期生存适应的结果,对物种生存繁衍具有重要意义。2017年4月至2019年7月,组在梵净山国家级自然保护区内布设了41台红外相机进行监测和研究。基于红外相机监测数据,利用占域模型（Occupancy model）和核密度估计方法（Kernel density estimation）,对食蟹獴（Herpestes urva）的栖息地选择和日活动节律进行了分析探讨。结果表明：（1）食蟹獴在常绿落叶阔叶混交林出现频率最高,在落叶阔叶林和针阔混交林出现频率最低,并集中出现于中低海拔区域;（2）在8种栖息地选择影响因素中,具有重要影响的因子为针阔混交林、海拔、水源地和常绿阔叶落叶混交林4种。其中,食蟹獴明显选择常绿阔叶落叶混交林（β=1.79±1.19 SE）,明显排斥针阔混交林（β=-1.93±1.30 SE）,海拔高度、离水源地的距离与占用率呈明显负相关;（3）年活动节律显示,食蟹獴在雨季活动明显增多,4-9月的活动频次在全年活动中占比达83.50%;（4）食蟹獴为完全昼行性物种,日活动时间分配较为平均,在9：00-10：00和13：00-14：00活跃度低,日活动高峰出现在11：00-13：00。梵净山地区作为食蟹獴分布区的北端,其典型的山地气候特征和植被类型与其他分布区存在显著差异,并面临不同的捕食风险和不同程度的人为干扰,食蟹獴对栖息地的选择方式和活动节律也出现了适应性的变化。     

深圳大鹏湾六种珊瑚共附生真菌多样性研究

本研究使用6种不同培养基对深圳大鹏湾6种珊瑚上的共附生真菌进行平板涂布法分离培养,结合ITS-rDNA基因序列特征和形态学特征进行鉴定来研究深圳大鹏湾海域珊瑚共附生真菌的多样性。共分离培养出457株真菌菌株,分属于7个属,分别为红酵母属Rhodotorula（180株）、曲霉属Aspergillus（170株）、木霉属Trichoderma（50株）、青霉属Penicillium（34株）、枝孢属Cladosporium（21株）、透孢黑团壳属Massarina（1株）和弯颈霉属Tolypocladium（1株）,其中优势属为红酵母属Rhodotorula和曲霉属Aspergillus,占菌株分离总数的76.59%。单独鹿角珊瑚上分离到的共附生真菌数量和种类最多,为160株9种;在该种珊瑚上还分离出弯颈霉属Tolypocladium sp.,该属真菌此前从未在珊瑚共附生真菌的研究中分离出来。本研究还发现SDA培养基分离出的共附生真菌数量最多,CDA培养基的真菌数量最少,表明不同培养基分离的共附生真菌多样性存在差异。     

转化异丁香酚生成香草醛纺锤芽孢杆菌的筛选

以底物异丁香酚为唯一碳源从七壤中筛选获得了一株能高效转化异丁香酚生成香草醛的芽孢杆菌。根据生理生化特性及16SrRNA序列分析鉴定其属于纺锤芽孢杆菌（Bacillus fusiformis）,初步试验表明该菌能转化2％异丁香酚生成4．20g/L香草醚。     

高产虾青素的红发夫酵母菌种的选育

红发夫酵母（Phaffia rhodozyma）是发酵法生产虾青素的优良菌株。本采用Cs^137-γ射线重复辐照,并交替进行亚硝基胍（NTG）诱变处理,选育得到一株高产虾青素的红夫酵母YB-20-29突变株。该菌株摇瓶发酵的生物量达36.32g/L,总色素含量为1216.0ug/g,较原始菌株提高308%,虾青素产量达30.9ug/mL,是一株很有开发前景和虾青素高产菌株。     

新型生物杀虫剂--刺糖菌素

刺糖菌素是由土壤放线菌多刺糖多孢菌（Saccharopolyspora spinosa）产生的次级代谢产物,是一种具有触杀及摄食毒性的广谱杀虫剂。对鳞翅目害虫而言,刺糖菌素是目前已发现的杀虫剂中选择性最高的化合物之一。中就刺糖菌素的结构、生物合成、性质以及生产方法进行了综述。     

水淹对三峡库区岸生植物秋华柳(Salix variegata Franch.)存活和恢复生长的影响

植物在水淹后的存活和恢复生长状况可以衡量其对水淹的耐受能力,为了解三峡库区岸生植物秋华柳(Salix variegata Franch.)的水淹耐受能力,研究了长期水淹条件下秋华柳植株的存活和恢复生长状况.实验设置了3个水淹深度:对照,水淹根部(植株置于水中,仅地下部分被淹没)和水淹2m(植株置于水中,顶部距水面2m),6个水淹时间:20d,40d,60d,90d,120d和180d.研究结果表明:(1)水淹对秋华柳植株存活率的影响较小.水淹根部处理的植株在水淹180d后,存活率仍为100%,水淹2m处理的植株在水淹120d后,存活率也为100%,直到180d后,存活率才下降为0.(2)水淹后,秋华柳植株仍然可以进行恢复生长,表现出很强的恢复生长能力,但因水淹处理的不同,其恢复生长存在差异.随着水淹时间的延长,秋华柳植株出水后到开始恢复生长之前所需的时间增加,但所有水淹处理的植株在水淹结束后一周内都可以开始恢复生长.在相同水淹时间处理下,水淹处理的秋华柳植株在恢复生长期间的相对生长速率都高于对照植株,水淹40d,60d,90d后,水淹2m的秋华柳植株分别比对照植株高57.8%,143.4%,130.4%.水淹结束时,秋华柳地上部分生物量随水淹深度的不同而不同,水淹根部处理的植株几乎与对照植株无显著差异,水淹2m处理的植株都低于对照植株.水淹结束后,不同处理的秋华柳植株生长2个月后的地上部分生物量与其在水淹结束时不同处理植株地上部分生物量的变化趋势相似.本研究表明,秋华柳在长时间的水淹后具有很高的存活率,并可以进行很好的恢复生长,表现出较强的水淹耐受能力,可以考虑将其应用于三峡库区消落区的植被构建.