水稻PHD-finger转录因子基因OsMsr16增强耐盐性的可能性研究

Os Msr16(Oryza sativa L.multi-stress-responsive gene 16)是一个新的水稻植物同源结构域(PHD)-finger家族转录因子基因。之前研究显示,Os Msr16受到低温、高温和干旱胁迫的诱导表达。本研究中,q RT-PCR分析表明Os Msr16同样受到盐胁迫的诱导表达,推测该基因可能参与植物在高盐胁迫下的生理调控。苗期盐处理条件下,与对照相比,过表达Os Msr16的转基因水稻植株叶片失绿面积较少,存活率更高。盐胁迫条件下,转基因植株中积累更高含量的脯氨酸和可溶性糖,而丙二醛含量和双氧水含量明显降低。转基因植株中超氧化物歧化酶(superoxide dismutase,SOD)活性和过氧化氢酶(catalase,CAT)活性明显高于对照。以上结果说明Os Msr16基因可能在植物防御盐胁迫过程中发挥重要作用。     

盐胁迫下OsDSR2 RNAi转基因水稻的生理特性及转录组学分析

为探究盐胁迫下OsDSR2 RNAi转基因水稻的生理特性和差异基因的表达调控,以中花11(ZH11)植株为对照,对OsDSR2 RNAi转基因水稻幼苗进行生理特性和转录组学分析,结果表明:正常条件下,ZH11和Os DSR2 RNAi转基因水稻中叶绿素含量和根部Na+/K+均显著低于ZH11,其他各项生理指标均没有显著性差异,而盐胁迫处理后,Os DSR2 RNAi转基因水稻中的叶绿素含量、可溶性糖(SS)含量、过氧化氢酶(CAT)活性与ZH11相比没有明显变化,但转基因植株中的细胞膜透性、丙二醛(MDA)含量、Na+/K+显著或极显著低于ZH11,脯氨酸(Pro)含量、超氧化物歧化酶(SOD)活性、过氧化物酶(POD)活性显著或极显著高于ZH11。Os DSR2 RNAi转基因水稻在盐胁迫前后共产生68个差异表达基因,其中有55个上调表达,13个下调表达。GO分析结果显示,差异表达基因主要富集在胁迫应激反应、分解代谢等生物学过程中。KEGG分析结果表明,差异表达基因主要参与类胡萝卜素生物合成、油菜素内酯生物合成以及表皮素,软木脂和蜡质生物合成代谢通路中,进一步通过RT-q PCR验证了Osb ZIP16、Os LEA3、RAB21等差异表达基因参与水稻的胁迫应答反应。综上,在生理层面上,Os DSR2 RNAi转基因水稻主要通过降低细胞膜透性、MDA含量和Na+/K+,增加Pro含量,提高SOD和POD活性,抑制叶绿素的降解来提高耐盐性;在分子水平上,Os DSR2主要通过脱落酸(ABA)、油菜素内酯(BR)信号通路参与调控Osb ZIP16、Os LEA3、RAB21等逆境相关基因的表达来提高苗期水稻的耐盐性,为进一步阐明Os DSR2参与调控水稻耐盐的详细分子机制奠定基础。     

异源表达AtCYSa基因烟草的耐盐和抗虫特性分析

植物半胱氨酸蛋白酶抑制剂在植物防御生物与非生物胁迫过程中发挥着重要的作用。拟南芥中的半胱氨酸蛋白酶抑制剂AtCYSa基因的表达能够受到多种胁迫的诱导,且在拟南芥中过量表达AtCYSa基因可以增强转基因拟南芥抵御盐、干旱和氧化等非生物胁迫的能力。为了进一步探究AtCYSa基因的功能以及在烟草中的应用,构建了植物表达载体pCAMBIA 1302-AtCYSa。通过PCR以及RT-PCR验证共获得4株阳性转基因烟草,选择其中3株转基因烟草进行盐胁迫处理和抗虫活性实验。在盐胁迫处理中,100mmol/L NaCl和200mmol/L NaCl处理组的丙二醛含量显著低于野生型对照组。伊文思蓝染色和细胞相对活性结果表明,转基因烟草的细胞活性比野生型烟草明显偏高。这说明在盐胁迫处理下,AtCYSa基因的表达能够起到保护转基因烟草的作用。抗虫活性研究发现,实验组的幼虫总重均呈明显的下降趋势,且幼虫死亡率显著高于对照组。这些结果表明,AtCYSa基因在烟草中的过量表达能够增强转基因烟草的耐盐以及抗虫的能力。     

一个水稻高效表达启动子Os252的分离

植物基因的表达受启动子的控制,高效表达启动子的分离及功能分析不仅是植物基因工程研究的重要研究方面,也是表达调控研究的重要内容。根据EST数据克隆了一个预测在水稻茎中高效表达的启动子Os252。将该启动子与GUS基因构建成表达载体并转入水稻。转基因水稻PCR分析表明,GUS基因已经成功地整合进水稻基因组中。GUS组织化学分析表明,Os252能启动GUS基因在水稻叶、茎以及胚乳中表达。进一步GUS酶活性的测定表明,叶和胚乳中Os252启动子活性分别是35S启动子的1.9和2.5倍。由于Os252来自于水稻,在叶和胚乳中活性高于35S启动子,因此该启动子可望用于水稻基因工程研究。     

水稻Bph14基因生理功能初探

Bph14基因是第一个被克隆的水稻抗虫基因,但鲜有Bph14基因对水稻生理方面影响的报道。研究该基因对水稻生理方面的影响有助于全面理解Bph14基因对褐飞虱的抗性机制。将Bph14基因转入水稻栽培品种中,对携带有外源基因的转基因株系通过荧光定量PCR检测乙烯合成和非生物胁迫相关基因的表达,另外,分别用5μmol/L ABA和250 mmol/L Na Cl处理转基因水稻株系和相应受体材料,结果表明,与乙烯合成相关的基因Os ACO2,Os ACS2,Os ACS6,O1g P5CS和O5g P5CS的表达量均较转基因受体m5274发生了变化,与干旱、耐盐等非生物胁迫相关的基因Cat A、Cat B、Cat C、RAB16A、LEA3、LIP9、Sal T、Adh I等也都不同于在受体m5274中的表达。同时Bph14基因的导入也提高了水稻对ABA的敏感性和对盐胁迫的耐受性,说明Bph14基因在水稻中可能参与调控乙烯合成以及非生物胁迫应答的功能基因的表达。     

转反义DsRACK1基因增强水稻抗旱能力

RAC1是一种多功能支架蛋白,广泛参与植物生长发育过程的调节。利用反义RNA技术抑制水稻（Oryzasativa）RACK1基因的表达,分析了RACK1基因在响应干旱胁迫中的功能。采用实时定量PCR对获得的转基因植株的RACK1基因表达进行分析,结果表明转基因水稻RACK1基因表达受抑制程度达到50％左右。与非转基因水稻（对照）相比,转基因水稻耐干旱能力强,膜脂过氧化程度低且丙二醛的含量少,SOD活性高。这些结果表明,RACK1蛋白负调节水稻对干旱胁迫的耐受过程,并且这种调节作用在很大程度上与植株体内的氧化还原系统有关。     

转反义OsRACK1 基因增强水稻抗旱能力

RACK1是一种多功能支架蛋白, 广泛参与植物生长发育过程的调节。利用反义RNA技术抑制水稻(Oryz a sativa)RACK1基因的表达, 分析了RACK1基因在响应干旱胁迫中的功能。采用实时定量PCR对获得的转基因植株的RACK1基因表达进行分析, 结果表明转基因水稻RACK1基因表达受抑制程度达到50%左右。与非转基因水稻(对照)相比, 转基因水稻耐干旱能力强, 膜脂过氧化程度低且丙二醛的含量少, SOD活性高。这些结果表明, RACK1蛋白负调节水稻对干旱胁迫的耐受过程, 并且这种调节作用在很大程度上与植株体内的氧化还原系统有关。     

转反义OsRACK1基因增强水稻抗旱能力

RAC1是一种多功能支架蛋白,广泛参与植物生长发育过程的调节。利用反义RNA技术抑制水稻（Oryzasativa）RACK1基因的表达,分析了RACK1基因在响应干旱胁迫中的功能。采用实时定量PCR对获得的转基因植株的RACK1基因表达进行分析,结果表明转基因水稻RACK1基因表达受抑制程度达到50％左右。与非转基因水稻（对照）相比,转基因水稻耐干旱能力强,膜脂过氧化程度低且丙二醛的含量少,SOD活性高。这些结果表明,RACK1蛋白负调节水稻对干旱胁迫的耐受过程,并且这种调节作用在很大程度上与植株体内的氧化还原系统有关。     

两个水稻胚乳启动子的克隆及表达分析

启动子的克隆对基因表达及基因工程研究有重要意义。根据数据库中EST丰度,从水稻中克隆了两个预测在水稻胚乳中高效表达的启动子Os772和Os359,并将启动子片段与GUS报告基因融合,构建了重组表达载体。通过农杆菌介导方法将其导入水稻愈伤组织细胞。转基因水稻经GUS组织化学分析显示,Os772和Os359能启动GUS基因在水稻胚乳中表达但不能在根、茎、叶和花中表达。该结果表明Os772和Os359为两个水稻胚乳特异性启动子。     

水稻基因Os922的克隆、转化及对葡萄糖的敏感性研究

浙江大学生物技术研究所白洋、郝中娜,中国农业大学刘东风、郭泽建等5位植物转基因科研工作者,通过PCR方法从水稻cDNA文库中克隆1个水稻AP2/EREBP类的转录因子基因Os922,并构建增强表达载体COU-Os922,使用光照培养方法将根癌农杆菌（Agrobacterium tume faciens）EH．ACOS转化到粳稻（Orgza sativa L．）秀水11的幼胚,得到转基因水稻植株。     

A Unique Protease Cleavage Site Predicted in the Spike Protein of the Novel Pneumonia Coronavirus (2019-nCoV) Potentially Related to Viral Transmissibility

正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases     

Curcuminoids as inhibitors of thioredoxin reductase: A receptor based pharmacophore study with distance mapping of the active site

Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable potent antiproliferative synthetic drugs.     

Comparative morphology of the carpel in the Liliaceae: Hewardieae, Petrosavieae, and Tricyrteae

The young pistils in the melanthioid tribes, Hewardieae, Petrosavieae and Tricyrteae, are uniformly tricarpellate and syncarpous. They lack raphide idioblasts. All are multiovulate, with bitegmic ovules. The Petrosavieae are marked by the presence of septal glands and incomplete syncarpy. Tepals and stamens adhere to the ovary in the Hewardieae and the Petrosavieae but not in the Tricyrteae. Two vascular bundles occur in the stamens of the Hewartlieae and Tricyrtis latifolia. Ventral bundles in the upper part of the ovary of the Hewardieae are continuous with compound septal bundles and placental bundles in the lower part. Putative ventral bundles occur in the alternate position in the Tricyrteae and putative placental bundles in the opposite. position in the Petrosavieae. The dichtomously branched stigma in each carpel of the Tricyrteae is supplied by a bifurcated dorsal bundle.     

Enterovirus 71 3C proteolytically processes the histone H3 N-terminal tail during infection

Highlights
1. The N-terminal tail of histone H3 is specifically cleaved during EV71 infection.
2. Viral protease 3C is identified as a protease responsible for proteolytically processing the N-terminal H3 tail.
3. Our finding reveals a new epigenetic regulatory mechanism for Enterovirus 71 in virus-host interactions.     

Detection of EBV and HHV6 in the Brain Tissue of Patients with Rasmussen’s Encephalitis

Rasmussen’s encephalitis (RE) is a rare pediatric neurological disorder, and the exact etiology is not clear. Viral infection may be involved in the pathogenesis of RE, but conflicting results have reported. In this study, we evaluated the expression of both Epstein-Barr virus (EBV) and human herpes virus (HHV) 6 antigens in brain sections from 30 patients with RE and 16 control individuals by immunohistochemistry. In the RE group, EBV and HHV6 antigens were detected in 56.7% (17/30) and 50% (15/30) of individuals, respectively. In contrast, no detectable EBV and HHV6 antigen expression was found in brain tissues of the control group. The co-expression of EBV and HHV6 was detected in 20.0% (6/30) of individuals. In particular, a 4-year-old boy had a typical clinical course, including a medical history of viral encephalitis, intractable epilepsy, and hemispheric atrophy. The co-expression of EBV and HHV6 was detected in neurons and astrocytes in the brain tissue, accompanied by a high frequency of CD8⁺ T cells. Our results suggest that EBV and HHV6 infection and the activation of CD8⁺ T cells are involved in the pathogenesis of RE.     

Perinatal Vertical Transmission of Chikungunya Virus in Ruili,a Town on the Border between China and Myanmar

正Dear Editor,Chikungunya virus (CHIKV), an arbovirus in the family of Togaviridae, genus Alphavirus, is transmitted by the A.aegyptii or A. albopictus mosquito, and causes disease in humans characterized by fever, rash, and arthralgia (Silva and Dermody 2017; Suhrbier 2019). It was first reported in 1953 in Tanzania, and caused only a few outbreaks and sporadic cases in Africa and Asia in last century. However, in the epidemic in 2004, CHIKV acquired mutations that conferred enhanced transmission by the A. albopictus mosquito(Schuffenecker et al. 2006). Since then, it has successively caused outbreaks in Africa, the Indian Ocean, South East Asia, the South America, and Europe (Zeller et al. 2016).     

Development of a Novel Reverse Transcription Loop-Mediated Isothermal Amplification Method for Rapid Detection of SARS-CoV-2

In conclusion, the novel visual RT-LAMP assay is a simple, rapid, and sensitive approach for detection of SARS-CoV-2, and it is ready for application in primary care and community hospitals or health care centers, and even patients' own houses in response to the current SARS-CoV-2 epidemic because the assay does not require sophisticated equipment and skilled personnel. Furthermore, it is also ready to be used in fields for screening samples from wild animals and environments to facilitate the identification of potential intermediate hosts that mediate the cross-species transmission of SARS-CoV-2 from bats to humans.