Expression of baculovirus anti-apoptotic genes p35 and op-iap in cotton (Gossypium hirsutum L.) enhances tolerance to verticillium wilt

Background

Programmed cell death plays an important role in mediating plant adaptive responses to the environment such as the invasion of pathogens. Verticillium wilt, caused by the necrotrophic pathogen Verticillium dahliae, is a serious vascular disease responsible for great economic losses to cotton, but the molecular mechanisms of verticillium disease and effective, safe methods of resistance to verticillium wilt remain unexplored.

Methodology/Principal Findings

In this study, we introduced baculovirus apoptosis inhibitor genes p35 and op-iap into the genome of cotton via Agrobacterium-mediated transformation and analyzed the response of transgenic plants to verticillium wilt. Results showed that p35 and op-iap constructs were stably integrated into the cotton genome, expressed in the transgenic lines, and inherited through the T₃ generation. The transgenic lines had significantly increased tolerance to verticillium wilt throughout the developmental stages. The disease index of T₁–T₃ generation was lower than 19, significantly (P<0.05) better than the negative control line z99668. After treatment with 250 mg/L VD-toxins for 36 hours, DNA from negative control leaves was fragmented, whereas fragmentation in the transgenic leaf DNA did not occur. The percentage of cell death in transgenic lines increased by 7.11% after 60 mg/L VD-toxin treatment, which was less than that of the negative control lines''s 21.27%. This indicates that p35 and op-iap gene expression partially protects cells from VD-toxin induced programmed cell death (PCD).

Conclusion/Significance

Verticillium dahliae can trigger plant cells to die through induction of a PCD mechanism involved in pathogenesis. This paper provides a potential strategy for engineering broad-spectrum necrotrophic disease resistance in plants.     

山东强筋和中筋小麦品质形成的气象条件及区划

1999—2003年选用4个强筋小麦品种和12个中筋小麦品种在山东省的31个县进行试验,以各试验点当地气象站观测的小麦开花至成熟期的气象资料为依据,采用逐步回归和一元非线性回归模型进行分析.结果表明:平均气温分别为20.0℃和20.5℃时,有利于强筋和中筋小麦面团稳定时间的延长;气温日较差分别为12.7℃和11.7℃时,有利于强筋和中筋小麦籽粒蛋白质含量的提高;降水量分别为48.6mm和52.1mm,日照时数分别为297h和299h时,有利于强筋和中筋小麦沉降值的提高;降水量分别为53.5mm和53.9mm,日照时数分别为295h和298h时,有利于强筋和中筋小麦面团稳定时间的延长.单一气象因子对小麦籽粒蛋白质含量、沉降值和面团稳定时间的影响不完全同步.依据对各地气象条件进行综合评价的结果,将山东省划分为胶东与鲁中强筋、中筋小麦适宜区,鲁西北和鲁西南强筋、中筋小麦次适宜区,鲁南强筋小麦次适宜、中筋小麦适宜区.     

A preliminary X-ray study of a refolded PTS EIIBfruc protein from Escherichia coli

The phosphoenolpyruvate-carbohydrate phosphotransferase system (PTS) catalyzes the phosphorylation and transportation of its sugar substrates. A sugar-specific enzyme II complex involved in the PTS finally functions to translocate substrates across the membrane. A PTS EIIB(fruc) protein, a fructose specific EIIB subunit, from Escherichia coli has been cloned, expressed, refolded, purified, and crystallized. The synchrotron data were collected to 2.6 A from the crystal of a selenomethionine substitute PTS EIIB(fruc) protein. The crystal belongs to the primitive trigonal space group P3(1)21, with unit-cell parameters of a = 33.4 A, b = 33.4 A, c = 154.0 A, and beta = 120.0 degrees . A full structure determination is under way to provide insights into the structure-function relationships of this protein.     

Imobilization of β-glucosidase using the cellulose-binding domain of Bacillus subtilis endo-β-1,4-glucanase

A recombinant plasmid pβCBD was constructed for immobilization of Cellulomonas fimi β-glucosidase (Cbg) using the cellulose-binding domain (CBD) of Bacillus subtilis BSE 616 endo-β-1,4-glucanase (Beg). The Cbg-CBD _Beg fusion protein, 80 kDa, was expressed in Escherichia coli and immobilized to Avicel. Cellobiose was completely hydrolyzed with the immobilized fusion protein. The fusion protein bound to Avicel retained full activity during continuous operation for 24 h at 4°C. This revised version was published online in November 2006 with corrections to the Cover Date.     

Crystal structure of type VI effector Tse1 from Pseudomonas aeruginosa

The type VI secretion systems (T6SS) have emerging roles in interspecies competition. In order to have an advantage in defense against other organisms, this system in Pseudomonas aeruginosa delivers a peptidoglycan amidase (Tse1) to the periplasmic space of a competitor. An immune protein (Tsi1) is also produced by the bacterium to protect itself from damage caused by Tse1. Tsi1 directly interacts with Tse1. We report that the crystal structure of Tse1 displays a common CHAP protein fold. Strikingly, our structures showed that the third residue in the catalytic triad may be novel as this residue type has not been observed previously.     

Induction of cytochrome P450 1A is required for circulation failure and edema by 2,3,7,8-tetrachlorodibenzo-p-dioxin in zebrafish

The mechanism of toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is thought to result from changes in gene expression via the aryl hydrocarbon receptor (AHR). The induction of cytochrome P450 1A (CYP1A) in various organs is a cardinal effect of TCDD. However, whether CYP1A is involved in endpoints of TCDD toxicity is controversial. We investigated the role of CYP1A in TCDD-induced developmental toxicities using gene knock-down with morpholino antisense oligos. Exposure of zebrafish embryos to TCDD, at concentrations eliciting the hallmark endpoints of developmental toxicity, induced CYP1A in the heart and vascular endothelium throughout the body. This induction by TCDD was markedly inhibited by morpholinos to zebrafish arylhydrocarbon receptor 2 (zfAHR2-MO) and to zebrafish CYP1A (zfCYP1A-MO). The zfAHR2-MO but not the zfCYP1A-MO inhibited zfCYP1A mRNA expression, indicating the specificities of these morpholinos. Injection of either zfAHR2-MO or zfCYP1A-MO blocked the representative signs of TCDD developmental toxicity in zebrafish, pericardial edema and trunk circulation failure. The morpholinos appeared do not affect normal development in TCDD-untreated embryos. These results suggest a mediatory role of zfCYP1A induction through zfAHR2 activation in causing circulation failure by TCDD in zebrafish. This is the first molecular evidence demonstrating an essential requirement for CYP1A induction in TCDD-evoked developmental toxicities in any vertebrate species.     

繁缕和无瓣繁缕六个居群的数值分析

对繁缕（Stellaria media）和无瓣繁缕（S.apetala）的6个居群的57个性状进行Q－聚类和R－聚类的研究。结果表明：（1）Q－聚类中,用一条结合线,可以把繁缕的4个居群聚为一类,无瓣繁缕的2个居群聚为一类。这一结果支持肥繁缕和无瓣繁缕划分为两个物种;（2）R－聚类中,发现了呈现完全正相关、极大正相关和极大负相关的性状,并根据R－聚类的结果,运用一条适当的结合线,把繁缕和无瓣繁缕的57个性状划为5个类群,并分析了各性状的分类学意义。     

A “GC-rich” method for mammalian gene expression: A dominant role of non-coding DNA GC content in regulation of mammalian gene expression

High mammalian gene expression was obtained for more than twenty different proteins in different cell types by just a few laboratory scale stable gene transfections for each protein. The stable expression vectors were constructed by inserting a naturally-occurring 1.006 kb or a synthetic 0.733 kb DNA fragment (including intron) of extremely GC-rich at the 5′ or/and 3′ flanking regions of these protein genes or their gene promoters. This experiment is the first experimental evidence showing that a non-coding extremely GC-rich DNA fragment is a super “chromatin opening element” and plays an important role in mammalian gene expression. This experiment has further indicated that chromatin-based regulation of mammalian gene expression is at least partially embedded in DNA primary structure, namely DNA GC-content.     

PERMANENT GENETIC RESOURCES: A set of primer pairs for amplifying the complete mitochondrial DNA of endangered Chinese egret (Aves, Ardeidae, Egretta eulophotes)

The Chinese egret is a globally endangered species. Here we describe a set of primer pairs to amplify its entire mtDNA. The polymerase chain reaction products (1000–2000 bp) were successfully amplified by using this primer set and were then sequenced and aligned. The contiguous mtDNA sequences of the Chinese egret were assembled to be a circular molecule (17 579 bp). This primer set was also confirmed to be useful for six other species of ardeid birds. The versatility of this primer set will provide a groundwork for further studies on the genetic structure and molecular evolution of the ardeid species.