Incidence and genetic diversity of raspberry bushy dwarf virus (RBDV) in Rubus spp. in Turkey

Raspberry bushy dwarf virus (RBDV), recently renamed to Idaeovirus rubi, is one of the most common viruses infecting Rubus species worldwide but there is still a limited number of genome sequences available in the GenBank database and the majority of the sequences include partial sequences of RNA-1 and RNA-2. The distribution and incidence of RBDV in main raspberry and blackberry growing provinces in Turkey were monitored during 2015–2019 and 537 Rubus spp. samples were tested by both DAS-ELISA and RT-PCR. Among the tested samples, 36 samples tested positive for RBDV by DAS-ELISA and 67 samples by RT-PCR. There was relatively low nucleotide diversity among the Turkish isolates. Turkish isolates shared 93%–97.7%, 84.3%–98.9%, and 85%–99.2% nucleotide sequence identities with available sequences in the GenBank, in partial RNA-1, movement protein (MP) and coat protein (CP) genes, respectively. In the phylogenetic tree constructed for RNA-1, MP, and CP sequences, all Turkish raspberry isolates were clustered in a distinct clade. However, the blackberry isolates showed considerable variation in nucleotide sequences and were placed in three distinct groups. The divergent blackberry isolates showed high variability in MP (84.5%–89.3%) and CP (85.5%–89.7%) regions and were placed in a distinct group. The rest of blackberry isolates clustered together with sweet cherry RBDV isolates adjacent to the grapevine clade or together with raspberry isolates. The comparative analysis conducted on three RNA segments of RBDV highlighted the high sequence diversity of Turkish RBDV isolates. This study also emphasizes the importance of regular monitoring of RBDV infections in Turkey, with special regard to those Rubus spp. and grapevine accessions employed in conservation and selection programmes. In particular, the presence of new RBDV genetic variants and infection of Rubus species must be taken into account to choose a correct detection protocol and management strategy.     

Rice ubiquitin-conjugating enzyme OsUbc13 negatively regulates immunity against pathogens by enhancing the activity of OsSnRK1a

Ubc13 is required for Lys63-linked polyubiquitination and innate immune responses in mammals, but its functions in plant immunity still remain largely unknown. Here, we used molecular biological, pathological, biochemical, and genetic approaches to evaluate the roles of rice OsUbc13 in response to pathogens. The OsUbc13-RNA interference (RNAi) lines with lesion mimic phenotypes displayed a significant increase in the accumulation of flg22- and chitin-induced reactive oxygen species, and in defence-related genes expression or hormones as well as resistance to Magnaporthe oryzae and Xanthomonas oryzae pv oryzae. Strikingly, OsUbc13 directly interacts with OsSnRK1a, which is the α catalytic subunit of SnRK1 (sucrose non-fermenting-1-related protein kinase-1) and acts as a positive regulator of broad-spectrum disease resistance in rice. In the OsUbc13-RNAi plants, although the protein level of OsSnRK1a did not change, its activity and ABA sensitivity were obviously enhanced, and the K63-linked polyubiquitination was weaker than that of wild-type Dongjin (DJ). Overexpression of the deubiquitinase-encoding gene OsOTUB1.1 produced similar effects with inhibition of OsUbc13 in affecting immunity responses, M. oryzae resistance, OsSnRK1a ubiquitination, and OsSnRK1a activity. Furthermore, re-interfering with OsSnRK1a in one OsUbc13-RNAi line (Ri-3) partially restored its M. oryzae resistance to a level between those of Ri-3 and DJ. Our data demonstrate OsUbc13 negatively regulates immunity against pathogens by enhancing the activity of OsSnRK1a.     

Responses of the desert green algae,Chlorella sp. to drought stress

Desert algae are important components of the desert soil crust and play an essential role in desert soil ecosystem development. Owing to their special habitat, desert algae are often exposed to harsh environments, among which drought represents the most common stress. Green algae are considered to have drought tolerance potential; however, only a few studies have investigated this. In this study, we selected the green alga Chlorella sp., which was isolated from desert soil, and studied its physiological response to polyethylene glycol (PEG) 6000-induced drought stress. The results showed that drought stress can affect the photosynthetic efficiency of Chlorella sp., reduce its water retention ability, and destroy its ultrastructure. However, Chlorella sp. can cope with drought stress through a series of physiological regulatory strategies. Protective strategies include quick recovery of photosynthetic efficiency and increased chlorophyll content. In addition, induced synthesis of soluble proteins, lipids, and extracellular polysaccharide (EPS), and accumulation of osmotic regulatory substances, such as sucrose and trehalose, also contribute to improving drought tolerance in Chlorella sp. This study provides insights into the physiological responses of Chlorella sp. to drought stress, which may be valuable for understanding the underlying drought adaptation mechanisms of desert green algae.     

Substitution of transferrin by FeCl3 in the development of a low foetal calf serum concentration medium for KB-26.5 hybridoma cell line

KB-26.5, a murine hybridoma cell line producing an IgG₃ monoclonal antibody used in blood type determination, primarily adapted to grow at 5% foetal calf serum (FCS) concentration has been adapted to grow at 0.5% FCS, maintaining its ability to produce antibodies at the same level. In the final step of adaptation, the addition of insulin, transferrin, ethanolamine and selenium to the media formulation was studied, using factorial assay techniques to check the effect of the different compounds and to optimize their required level for satisfactory growth and antibody secretion. KB-26.5 cells required only 20 g/ml of transferrin to adapt to 0.5% FCS medium. Furthermore, transferrin could be substituted by FeCl₃, at a relatively low level of 2 g/ml. Maximum cell density decreased by 31.5% in spinner flask test, but the antibody titer was maintained, thus the specific productivity increased. However, inoculum size had to be increased three-fold with 0.5% FCS medium in order to assure cell growth.     

湖南郴州发现岭南两头蛇

2022年8月,于湖南省郴州市永兴县湘阴渡镇（113°01′18′′E,26°11′87′′N,海拔174 m）采集到两头蛇活体标本1号,通过形态测量比较,发现所采集标本与中国广东省分布的岭南两头蛇（Calamariaarcana）形态特征基本一致;后基于线粒体Cytb基因片段进行分子系统发育分析,结果显示该标本与岭南两头蛇形成高支持率单系群（后验概率PP为1.00,自举值BS为100）,所采集标本与岭南两头蛇的遗传距离为1%。综合形态学和分子生物学证据,最终确定该标本为两头蛇科（Calamariidae）两头蛇属的岭南两头蛇,系湖南省蛇类分布新记录种。     

Comprehensive Evaluation and Screening for 1,2,4-Trichlorobenzene Tolerance in Rice Cultivars at Different Growth Stages

Journal of Plant Growth Regulation - 1,2,4-trichlorobenzene (1,2,4-TCB) pollution in fields has become a potential threat to rice growth, yet little is known about the different response of many...     

代谢工程改造大肠杆菌一碳模块高效合成L-甲硫氨酸

L-甲硫氨酸又名L-蛋氨酸,是人体必需8种氨基酸之一,在饲料、医药、食品领域具有重要应用。以实验室前期构建的M2(Escherichia coli W3110?IJAHFEBC/PAM)为出发菌株,以模块化代谢工程策略构建了一株L-甲硫氨酸高产菌株。首先通过过表达亚甲基四氢叶酸还原酶(methylenetetrahydrofolate reductase,MetF)和筛选不同来源的丝氨酸羟甲基转移酶(hydroxymethyltransferase,GlyA),增强了一碳模块甲基供体的生成,优化了一碳模块。随后针对一碳模块的前体供应,过表达了胱醚裂解酶(cysteamine lyase,MalY)和半胱氨酸内运基因(fliY),有效地提高了L-高半胱氨酸和L-半胱氨酸的供应。最终摇瓶发酵L-甲硫氨酸的产量由2.8 g/L提高至4.05 g/L,5 L发酵罐中达到18.26 g/L。研究结果表明,一碳模块对L-甲硫氨酸的生物合成具有十分重要的影响,在细胞内通过优化一碳模块,可以实现L-甲硫氨酸的高效生物合成。本研究为进一步提高微生物发酵生产L-甲硫氨酸的水平奠定了基础。     

光枝勾儿茶内生真菌多样性及抑菌活性研究

为探索贵州苗药光枝勾儿茶内生真菌类群特征、分布部位及其抑菌活性,该研究采用传统方法对贵州省贵阳市和黔西市光枝勾儿茶内生真菌进行分离,并基于分子生物学及统计学对其分类地位进行鉴定及多样性评价,最后通过微量肉汤倍比稀释法筛选具有抑菌活性的菌株。结果表明:(1)从光枝勾儿茶中分离到191 株内生真菌,隶属于3 个门5 个纲10 个目15 个科19 个属,优势属为叶点霉属(Phyllosticta)、间座壳属(Diaporthe)、葡萄座腔菌属(Botryosphaeria)和刺盘孢属(Colletotrichum)。(2)黔西光枝勾儿茶内生真菌香农-维纳多样性指数(H''_Q=2.112)较贵阳(H''_G=1.801)高,索伦森相似性指数Cs_G-Q为0.923,不同组织香农-维纳多样性指数为茎(H''_S=2.004)>根(H''_R=1.764)>叶(H''_L=1.654)>果实(H''_F=1.473),茎和叶内生真菌的索伦森相似性最高(Cs_S-L=0.667)。(3)筛选出的21株内生真菌对供试菌大肠杆菌(Escherichia coli)、金黄色葡萄球菌(Staphylococcus aureus)和沙门氏菌(Salmonella enterica)具有抑菌效果,其中Diaporthe sp. QX4G6对大肠杆菌、金黄色葡萄球菌和沙门氏菌的最小抑菌浓度分别为12.5、6.25、12.5 mg·mL^-1,最小杀菌浓度分别为12.5、6.25、12.5 mg·mL^-1。以上研究结果揭示了光枝勾儿茶蕴藏丰富的内生真菌资源,不同地区及组织内生真菌类群组成有差异,多个分离菌株具有抗菌活性,为光枝勾儿茶内生真菌天然抗菌药物或药源研发奠定了基础。     

云南省耿马佛洞地遗址发掘简报

佛洞地遗址位于云南省临沧市耿马傣族佤族自治县勐简乡勐简村大军赛村民小组燕子洞,坐落于一处东南开口的二叠纪灰岩穿洞,南临南汀河。2016～2017年,临沧市文物管理所在公路考古调勘期间发现该遗址;为进一步认识滇西地区旧石器时代晚期文化,2017～2018年对该遗址开展考古发掘工作。发掘区域位于洞内第四台面到第五台面间,共发掘20 m²,出土了包括石制品、动植物化石等在内的大量遗物。初步地层年代学分析显示,遗址时代为距今18400～14000年,共包含3期连续文化,文化遗物以石制品为主,总数达到9735件。佛洞地遗址作为一处热带-亚热带生境下的史前遗址,为我们构建旧石器时代晚期滇西地区文化序列、探讨特定自然生态背景下史前人类的文化适应提供了重要参考。