光源或培养基成分对金花茶愈伤组织中DFR、LAR与PPO基因表达及总儿茶素含量的影响

该研究以富含儿茶素的金花茶愈伤组织为材料,对不同光源、激素、碳源及苯丙氨酸处理30 d的愈伤组织中DFR表达量、LAR表达量、PPO表达量与总儿茶素含量的变化情况及四者两两之间的相关性进行了分析.结果表明:这4个检测项目均对以上处理有显著的响应;在以上各因素处理下,DFR与LAR的表达模式十分相似,其相关系数处在0.710~0.889之间;在不同碳源处理下,PPO表达量与总儿茶素含量的变化呈显著负相关关系,其相关系数为-0.696;在不同苯丙氨酸添加量处理下,DFR与LAR表达量变化均与总儿茶素含量变化呈显著正相关关系,其相关系数分别为0.786和0.564;适宜儿茶素离体生产的金花茶愈伤组织增殖配方为附加4 mg·L-16-BA、0.6 mg·L-12,4-D、30 g·L-1蔗糖与0.6608 g·L-1苯丙氨酸的MS固体培养基,其总儿茶素含量可达40.11 mg·g-1 DW.以上研究表明,与茶树相似,在金花茶中DFR与LAR在儿茶素代谢过程中密切相关;PPO表达量升高导致金花茶儿茶素损失;添加适宜浓度的苯丙氨酸作为前体物质是提高愈伤组织中总儿茶素含量的有效措施.     

锁阳愈伤组织诱导和增殖及不定根分化

以药用寄生植物锁阳的不同部位肉质茎为外植体,研究外植体形态及植物生长调节剂配比对愈伤组织形成、增殖及不定根分化的影响,建立了高效的锁阳肉质茎愈伤组织诱导、增殖和不定根分化体系。结果表明,锁阳茎下部大小为1．5cmx1．5cmxl．5cm的外植体,维管束平行于培养基放置,有利于愈伤组织形成;外植体培养50d,愈伤组织形成。高效的愈伤组织诱导培养基为Ms＋6-BA1．0mg.L-1＋2,4-D3．0mg·L-1,愈伤组织诱导率可达67％;增殖培养基为Ms＋6．BA0．5mg·L-1。＋2,4-D1．5mg·L-1,NxsN74％;在Ms＋6．BA1．0mg·L-1＋NAA2．0mg·L-1。分化培养基中,不定根诱导率达56％。     

海南粗榧愈伤组织的诱导和培养

海南粗榧的嫩茎和嫩叶以0.1%升汞消毒12 min的效果最好.外植体在MS 0.1 mg·L.6-BA l mg·L-1NAA 2mg·L-12,4-D 3.0%蔗糖 0.6%卡拉胶(pH 5.8)培养基中能顺利诱导出愈伤组织.液体悬浮和暗培养均有利于愈伤组织的增殖.愈伤组织增殖的最佳培养基为MS 0.1 mg·L-1KT 3 mg·L-1NAA 3.0%蔗糖(pH 5.8).     

不同抗生素对金发草愈伤组织生长分化的影响

该研究以金发草愈伤组织为材料,通过分析比较不同抗生素种类(卡那霉素、潮霉素、头孢噻呋钠和氨苄青霉素)和浓度对金发草愈伤组织生长分化的影响,来确定适用于金发草遗传转化体系中的抗性筛选剂和抑菌剂。结果表明：(1)金发草愈伤组织对卡那霉素很敏感,且其分化率随着卡那霉素浓度的增加显著减少( P＝0.01)。当卡那霉素浓度为10 mg·L-1时,金发草愈伤组织的生长分化受到明显抑制,且有大量的白化苗形成,但分化率仍有36.56％;当卡那霉素浓度为15 mg·L-1时,金发草愈伤组织的分化率为11.94％,只有很少部分的愈伤分化出绿色的丛生苗;当卡那霉素浓度为20 mg·L-1时,金发草愈伤组织基本褐化死亡,分化率仅为2.26％。因此,浓度为15 mg·L-1的卡那霉素适合作为金发草遗传转化体系中的抗性筛选剂。(2)金发草愈伤组织对潮霉素的敏感性要比卡那霉素弱,且潮霉素对金发草愈伤组织分化率的影响小,但毒害作用大。因此,潮霉素不适合作为金发草遗传转化体系中的抗性筛选剂。(3)300 mg·L-1的头孢霉素和氨苄青霉素对金发草愈伤组织生长分化影响很小且能有效抑制杂菌的生长,较高浓度的氨苄青霉素对金发草愈伤组织的抑制作用不太明显。因此,300 mg·L-1的头孢霉素和较高浓度的氨苄青霉素均可作为金发草遗传转化体系中的抑菌剂。该研究确定了适用于农杆菌介导的金发草遗传转化体系中的抗性筛选剂和抑菌剂,为金发草的遗传改良及功能性基因的研究奠定了基础。     

两个薄荷品种叶片愈伤组织的诱导及增殖

采用正交实验设计,研究了诱导培养基中6-BA、NAA和VC浓度及基本培养基类型4个因素对薄荷(Mentha haplocalyx Briq. )品种'687'和'沪39'叶片愈伤组织诱导的影响, 并研究了增殖培养基中6-BA、 NAA、 IBA和 2,4-D浓度对愈伤组织增殖的影响,筛选出适宜于品种'687'和'沪39'叶片愈伤组织诱导和增殖的最适培养基.结果表明,培养基中6-BA、NAA和VC浓度及基本培养基类型对品种'687'和'沪39'叶片愈伤组织的诱导率和褐化率均有一定的影响,但不同品种适宜的培养基配比有一定的差异;NAA浓度对品种'687'愈伤组织诱导的影响作用显著,影响品种'沪39'愈伤组织诱导的主要因素是VC浓度和基本培养基类型.适宜于品种'687'叶片愈伤组织诱导的培养基为含0.5 mg·L-1 6-BA和1.5 mg·L-1 NAA的B5培养基,适宜于品种'沪39' 愈伤组织诱导的培养基为含 1.5 mg·L-1 6-BA、 1.5 mg·L-1 NAA 和 100 mg·L-1 VC 的 1/2MS 培养基, 均含 5.5 g·L-1 琼脂和 30 g·L-1蔗糖(pH 5.5～pH 5.8).2个品种叶片愈伤组织增殖所需的生长调节剂浓度及比例也有一定的差异,较高浓度的细胞分裂素有利于愈伤组织的增殖,品种'687' 和'沪39'叶片愈伤组织增殖的适宜培养基分别为B5培养基和1/2MS培养基,均含2.0 mg·L-1 6-BA和1.5 mg·L-1 NAA以及5.5 g·L-1琼脂和30 g·L-1蔗糖(pH 5.5～pH 5.8).     

外源激素对杜衡叶柄愈伤组织诱导与分化的影响

以杜衡(Asarum forbesii Maxim.)叶柄为外植体,采用L9(34)正交实验设计分别研究了培养基中外源激素的种类及质量浓度对杜衡叶柄愈伤组织诱导和分化的影响,并据此筛选出适宜的诱导及分化培养基.结果表明:在杜衡叶柄愈伤组织的诱导过程中,培养基中细胞分裂素的种类(1.00 mg·L-16-BA、1.00 mg·L-1KT和1.00 mg·L-1ZT)和NAA质量浓度(0.00、0.10和0.30 mg·L-1)的影响效应均不显著,而2,4-D质量浓度(0.10、0.50和1.00 mg·L-1)则有显著影响(P<0.05);在愈伤组织的分化培养过程中,NAA(0.10、0.30和0.50 mg·L-1)的影响效应大于6-BA(1.00、3.00和5.00 mg·L-1)和IBA(0.01、0.05和0.10 mg·L-1).综合比较结果显示,适宜于杜衡叶柄愈伤组织诱导的培养基为添加1.00 mg·L-16-BA、0.30 mg·L-1NAA和1.00 mg·L-12,4-D的MS培养基(含6.5 g·L-1琼脂和30 g·L-1蔗糖,pH 5.8～pH 6.0),在此培养基上愈伤组织诱导率达到83.33%,且愈伤组织生长速度快、颗粒紧密;适宜于杜衡愈伤组织分化和不定芽增殖的培养基为添加3.00 mg·L-16-BA、0.10 mg·L-1IBA和0.30 mg·L-1NAA的MS培养基(含6.5 g·L-1琼脂和30 g·L-1蔗糖,pH 5.8～pH 6.0),在此培养基上愈伤组织的分化率最高(达到53.33%),增殖系数也最高(3.13).     

苦荞胚性愈伤组织诱导与植株再生研究

以苦荞子叶和下胚轴为外植体,进行了不同浓度激素组合的MS和SH固体培养基对胚性愈伤组织诱导及植株再生的研究。结果发现,MS培养基比SH培养基更有利于胚性愈伤组织诱导;2,4-D是诱导愈伤组织的有效激素,KT能有效促进胚状体的形成;下胚轴和子叶都能有效诱导出胚性愈伤组织和再生植株。下胚轴在MS 1.5mg·L-12,4-D 1.5mg·L-1BA培养基,子叶在MS 2mg·L-12,4-D 0.5~1.5mg·L-1BA上能高效诱导出愈伤组织;愈伤组织在MS 2mg·L-12,4-D 0.1mg·L-1KT培养基中继代,能有效诱导胚性愈伤组织;来自下胚轴的胚性愈伤组织在1/2MS 2.0mg·L-1BA 0.5mg·L-1KT 0.1mg·L-1NAA培养基上能够高频再生出芽,来自子叶的胚性愈伤组织在1/2MS 1.0mg·L-1BA 0.1mg·L-1KT 0.1mg·L-1NAA培养基上芽诱导率较高;MS 1mg·L-1NAA是适宜的再生苗生根培养基。     

甘薯叶柄原生质体有效植株再生

将甘薯(Ipomoea batatas (L．)Lam．)‘元气’和‘白星’(‘White Star’)的叶柄原生质体培养在含有0.05 mg·L-1 2,4-D和0.5 mg·L-1 KT的改良MS液体培养基中,3～4 d后细胞开始分裂。培养8～9周后,将直径达1～2 mm的愈伤组织转移到添加0.05～0.2 mg· L-1 2,4-D和0～0.5 mg·L-1 KT或添加0.5～2.0 mg ·L-1 NAA和1.0～3.0 mg·L-1 BAP 的MS固体增殖培养基上使其增殖。转移3～5周后,将愈伤组织再转移到MS基本培养基或转移到添加2.0～3.0 mg·L-1 BAP的MS培养基上。当进一步转移到MS基本培养基上后,从愈伤组织或从愈伤组织形成的不定根上再生出植株。‘元气’植株再生率高达60.0 ％,White Star高达43.4％。     

文心兰类原球茎的愈伤组织诱导及其植株再生

该研究首次以文心兰的类原球茎( protocorm-like bodies, PLBs)为外植体进行愈伤组织诱导及其植株再生培养,并分析了不同浓度的TDZ和2,4-D配比对愈伤组织增殖的影响。结果表明：以1/2MS为基本培养基,添加1 mg.L-1 TDZ与3 mg.L-12,4-D,从接种的 PLBs上可以诱导出乳白色的、较疏松的愈伤组织,诱导频率达到100％。愈伤组织继代培养时,在2,4-D浓度为0.5~2.0 mg.L-1的范围内,其增殖主要受TDZ浓度的影响,TDZ浓度从1.0 mg.L-1降低到0.5 mg.L-1,愈伤组织鲜重增殖倍数显著增加,由最低的4.50倍增加到最高的6.04倍。愈伤组织增殖的最适培养基为1/2MS ＋0.5 mg.L-1 TDZ ＋1.0 mg.L-12,4-D。将在最适愈伤组织增殖培养基上继代培养约1个月的愈伤组织转移到T2培养基(3.5 g.L-1花宝1号＋20 g.L-1红薯＋25 g.L-1香蕉＋1 g.L-1 tryptone ＋20 g.L-1蔗糖＋3.5 g.L-1 phytagel)上,黑暗培养1个月后,每克鲜重的愈伤组织约诱导出1328.67个PLBs。将诱导出的PLBs转移到新鲜的T2培养基上光照培养1个月,萌发率为90.12％。而将小植株转移到添加1 g.L-1活性炭的1/2MS培养基上,成苗率达到100％。该研究结果成功建立了文心兰的高频愈伤组织诱导及其植株再生体系,为文心兰基因工程育种提供了一个高效、稳定的转化受体系统。     

马尾松胚乳愈伤组织发生的初步研究

为获取马尾松(Pinus massoniana)单倍体材料，建立其胚乳组织培养体系，对马尾松胚乳不同的消毒方法进行筛选，并分析不同基本培养基、生长调节剂配比和热激处理对愈伤诱导的影响。结果表明，胚乳的最佳消毒方式为75%酒精处理30 s+2% NaClO消毒15 min，外植体污染率为0，死亡率仅为15.56%；生长素可促进愈伤诱导且是必需的；细胞分裂素浸泡种子可代替培养基中的细胞分裂素，并能取得比较好的胚乳愈伤诱导效果，1.0 mg·mL-1 6-BA浸泡30 min处理的效果最佳，在培养基WPM+NAA 2.0 mg·L-1+2,4-D 1.0 mg·L-1+蔗糖30 g·L-1+琼脂7 g·L-1中，愈伤诱导率高达87.78%；培养基DCR+NAA 2.0 mg·L-1+6-BA 1.0 mg·L-1+蔗糖30 g·L-1+琼脂7 g·L-1的胚乳愈伤诱导率最高(68.75%)；热激处理可提高胚乳愈伤的诱导率和质量，以相对湿度85%下45 ℃热激10 min最佳，总愈伤诱导率为66.25%，成团愈伤诱导率达8.75%；增殖培养以培养基WPM+NAA 0.1 mg·L-1+6-BA 2.0 mg·L-1+蔗糖30 g·L-1+琼脂7 g·L-1最佳，增殖率达83.33%。     

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.