Molecular Characterization of Cotton 14-3-3L Gene Preferentially Expressed During Fiber Elongation

The 14-3-3 protein, highly conserved in all eukaryotic cells, is an important regulatory protein. It plays an important role in the growth, amplification, apoptosis, signal transduction, and other crucial life activities of cells. A eDNA encoding a putative 14-3-3 protein was isolated from cotton fiber eDNA library. The eDNA, designated as Gh14-3-3L （Gossypium hirsutum 14-3-3-like）, is 1,029 bp in length （including a 762 bp long open reading frame and 5＇-/3＇-untranslated regions） and deduced a protein with 253 amino acids. The GhI4-3-3L shares higher homology with the known plant 14-3-3 proteins, and possesses the basic structure of 14-3-3 proteins： one dimeric domain, one phosphoralated-serine rich motif, four CC domains, and one EF Hand motif. Northern blotting analysis showed that Gh14-3-3L was predominantly expressed during early fiber development, and reached to the peak of expression in 10 days post anthers （DPA） fiber cells, suggesting that the gene may be involved in regulating fiber elongation. The gene is also expressed at higher level in both ovule and petal, but displays lower or undeteetable level of activity in other tissues of cotton.

Expression Analysis of 14-3-3 Gene in Tall Fescue under Several Abiotic Stresses

To study the functions of 14-3-3 gene family in tall fescue, the potential functions of 13 14-3-3 proteins in Arabidopsis were investigated by bioinformatic analysis. Based on the sequences of 14-3-3 genes in tall fescue by transcriptome and proteomic sequencing, the full-length cDNA sequences of 4 14-3-3 genes in tall fescue were obtained. Their sequences were aligned by Clustal W2. The results showed that the genetic relationships between 14-3-3A and 14-3-3D, 14-3-3B and 14-3-3C are closer, and their main structures are very conservative. The changes in expression levels of 14-3-3 genes under low nitrogen, drought, high temperature and high salt stresses were investigated by fluorescence quantitative PCR. The expres- sion level of 14-3-3A makes responses to low nitrogen, drought, high temperature and high salt stresses; the expression levels of other genes also make responses to abiotic stresses in varying degrees, but the relevant response mechanisms are not exactly the same. Therefore, it is speculated that the 14-3-3 gene family regu- lates stress resistance of plants through different pathways, and functional differenti- ation occurs during its evolution.

拟南芥FD与14-3-3/GRF7蛋白的相互作用研究

为了明确拟南芥FD与14-3-3/GRFs家族成员14-3-3/GRF7之间的互作关系,利用酵母双杂交和双分子荧光互补两种技术分别做了研究。在酵母中,与阴性对照相比,共转化FD-BD与14-3-3/GRF7-AD的酵母菌落在-Leu/-Trp/-His/-Ade四缺培养基上生长良好,表明在酵母中FD与4-3-3/GRF7之间存在直接的相互作用。在烟草中,与阴性对照相比,转化FD-c YFP与14-3-3/GRF7-n YFP,以及FD-n YFP与14-3-3/GRF7-c YFP载体的农杆菌共注射烟草细胞之后均在细胞核内观察到很强的荧光信号,表明在烟草细胞中FD与14-3-3/GRF7之间存在直接的相互作用。两种技术的结果共同证实FD与14-3-3/GRFs家族成员14-3-3/GRF7之间存在直接相互作用。

Wheat 14-3-3 Protein Conferring Growth Retardation in Arabidopsis

14-3-3 proteins belong to a family of phosphoserine/threonine-binding modules and participate in a wide array of signal transduction and regulatory events. Our previous study demonstrated that Ta14-3-3 was significantly down-regulated in leaf and root tissues of hybrid wheat at the tillering stage. In this paper, three homoeologous Ta14-3-3 genes were cloned from common wheat （Triticum aestivum L., 2n=6x=42, AABBDD） and mapped on chromosomes 2A, 2B, and 2D, respectively. Transgenic Arabidopsis plants ectopically overexpressing Ta14-3-3 displayed shorter primary roots, delayed flowering and retarded growth rates, indicating that Ta14-3-3 acted as a growth inhibitor in Arabidopsis. In wheat, Ta14-3-3 was down-regulated in roots and leaves of hybrids as compared to their parental lines. We proposed that Ta14-3-3 proteins might regulate growth vigor in hybrid wheat.

Creutzfeldt-Jakob disease presenting as Korsakoff syndrome caused by E196A mutation in PRNP gene:A case report

BACKGROUND Prion diseases are a group of degenerative nerve diseases that are caused by infectious prion proteins or gene mutations.In humans,prion diseases result from mutations in the prion protein gene(PRNP).Only a limited number of cases involving a specific PRNP mutation at codon 196(E196A)have been reported.The coexistence of Korsakoff syndrome in patients with Creutzfeldt-Jakob disease(CJD)caused by E196A mutation has not been documented in the existing literature.CASE SUMMARY A 61-year-old Chinese man initially presented with Korsakoff syndrome,followed by rapid-onset dementia,visual hallucinations,akinetic mutism,myoclonus,and hyperthermia.The patient had no significant personal or familial medical history.Magnetic resonance imaging of the brain revealed extensive hyperintense signals in the cortex,while positron emission tomography/computed tomography showed a diffuse reduction in cerebral cortex metabolism.Routine biochemical and microorganism testing of the cerebrospinal fluid(CSF)yielded normal results.Tests for thyroid function,human immunodeficiency virus,syphilis,vitamin B1 and B12 levels,and autoimmune rheumatic disorders were normal.Blood and CSF tests for autoimmune encephalitis and autoantibody-associated paraneoplastic syndrome yielded negative results.A test for 14-3-3 protein in the CSF yielded negative results.Whole-genome sequencing revealed a diseasecausing mutation in PRNP.The patient succumbed to the illness 11 months after the initial symptom onset.CONCLUSION Korsakoff syndrome,typically associated with alcohol intoxication,also manifests in CJD patients.Individuals with CJD along with PRNP E196A mutation may present with Korsakoff syndrome.

葫芦科14-3-3(GRF)基因家族的鉴定及进化分析

14-3-3蛋白是植物中一类高度保守的蛋白家族,因其特异识别并结合靶蛋白的磷酸化位点而广泛参与植物生长发育和逆境胁迫响应过程。为系统地了解14-3-3(GRF)基因家族在葫芦科作物中的基本特征和进化关系,本研究利用生物信息学从葫芦科7个物种基因组内共鉴定出83个14-3-3(GRF)基因,其中西瓜中有10个、甜瓜中9个、黄瓜中10个、瓠瓜中9个,3个南瓜属物种基因组各15个。理化特性表明14-3-3(GRF)蛋白的分子量约为30 k D,且大都为酸性氨基酸(pI<7.0)。相对于西瓜、甜瓜、黄瓜和瓠瓜,基因组复制事件是3个南瓜属物种内14-3-3(GRF)基因数目增多的主要原因。进化分析显示,葫芦科14-3-3(GRF)基因家族可进一步被划分为ε类和非ε类亚组,且后者的基因结构(外显子数目,内含子相位)较为保守。本研究结果首次完成了14-3-3(GRF)基因家族在葫芦科作物的鉴定、基因结构、基因家族扩增、共线性及进化分析,为更深入研究该家族基因的生物学功能提供参考。

辣椒14-3-3蛋白基因家族全基因组鉴定与表达特征分析

14-3-3蛋白(GRF)基因家族通过与靶蛋白相互作用,广泛参与植物激素信号转导和生理代谢过程,在植株生长发育和逆境响应中发挥重要作用。本研究以辣椒(Capsicum annuum)为材料,利用生物信息学研究方法,鉴定得到15个辣椒14-3-3基因家族成员,命名为CaGRF1~CaGRF15。系统分析了CaGRF基因家族成员的结构、进化关系、启动子顺式作用元件、组织及冷胁迫表达,并对CaGRF蛋白互作网络进行预测。结果表明,根据进化关系CaGRF家族成员被分为ε类和非ε类,ε类CaGRF含有更多外显子和内含子;启动子序列分析发现CaGRF启动子中有多个响应激素、胁迫、光的顺式作用元件;表达分析表明CaGRF各成员在辣椒各组织和冷胁迫响应中特异性表达;蛋白互作网络预测发现CaGRF可能与氮代谢、质子转运、转录调控相关蛋白互作。本研究为深入研究辣椒14-3-3家族成员功能及调控途径提供了参考。

超量表达GRF9基因促进番茄生长并增强其对磷的吸收利用能力

研究了编码拟南芥14-3-3蛋白的基因grf9导入番茄并超量表达后,转grf9基因番茄在土壤不同供磷条件下的生长和对磷的利用特征。结果发现,在2个供磷水平下,转grf9基因番茄的地上部含磷量均显著高于野生型,且分别提高58.70%和56.68%,磷素累积量较野生型增加88%～90%;同时转grf9基因番茄的地上部干重较野生型增加幅度约20%。高供磷条件下种植转grf9基因番茄的土壤速效磷耗竭程度显著高于种植野生型番茄的土壤,亏缺度增加15.7%;相应地,高供磷条件下转基因番茄对土壤磷的农学利用率和生理利用率均有显著提高,分别增强达200%和165%。该结果为培育磷高效新品种提供了新思路和科学依据。