Transgenic Indian mustard (<Emphasis Type="Italic">Brassica juncea</Emphasis>) expressing tomato glucanase leads to arrested growth of <Emphasis Type="Italic">Alternaria brassicae</Emphasis>

Brassica juncea is an important oilseed crop of the Indian sub-continent. Yield loss due to fungal disease alternaria leaf spot caused by Alternaria brassicae is a serious problem in cultivation of this crop. Nonavailability of resistance genes within crossable germplasms of Brassica necessitates use of genetic engineering strategies to develop genetic resistance against this pathogen. The pathogenesis related (PR) proteins are group of plant proteins that are toxic to invading fungal pathogens, but are present in plant in trace amount. Thus, overexpression of PR proteins leads to increased resistance to pathogenic fungi in several crops. The PR protein glucanase hydrolyzes a major cell-wall component, glucan, of pathogenic fungi and acts as a plant defense barrier. We report the expression of a class I basic glucanase gene, under the control of CaMV 35S promoter, in Indian mustard and its genetic resistance against alternaria leaf spot. Southern and Northern hybridization confirmed stable integration and expression of the glucanase gene in mustard transgenics. Several independent transgenics were screened in vitro and under poly house conditions for their resistance against Alternaria brassicae. In an in vitro antifungal assay, transgenics arrested hyphal growth of Alternaria brassicae by 15-54%. Under pathogen-challenged conditions in poly house, the transgenics showed restricted number, size and spread of lesions caused by Alternaria brassicae. Also, the onset of disease was delayed in transgenics compared to untransformed parent plants. The results demonstrate potentiality of a PR protein from a heterologous source in developing alternaria leaf spot resistance in Indian mustard.     

Effect of crop rotation on native vesicular arbuscular mycorrhizal propagules in soil

The effect of crop rotation of native vesicular arbuscular mycorrhizal fungi was studied. Finger millet was grown as the first season crop in 15 plots. In the second season a mycorhizal host (cowpea) and a non-mycorrhizal host (mustard) were grown in 5 plots each, and the remaining 5 plots were left fallow. In the third season cowpea was grown in all the plots. Leaving the land fallow reduced the mycorrhizal propagules by 40% while growing a non-mycorrhizal host reduced it by 13%. Cowpea grown in the third season coincided with a slow build up of mycorrhizal propagules in soil. There was a slow build up of mycorrhizal propagules late in the season irrespective of the treatment in the preceding season.     

Prototyping kinase inhibitor-cytotoxin anticancer mutual prodrugs activated by tumour hypoxia: A chemical proof of concept study

Amide- and ester-linked kinase inhibitor-cytotoxin conjugates were rationally designed and synthesised as prototype hypoxia-activated anticancer mutual prodrugs. Chemical reduction of an aryl nitro trigger moiety was shown to initiate a spontaneous cyclisation/fragmentation reaction that simultaneously released the kinase inhibitor semaxanib (SU5416) and the amine- or alcohol-linked cytotoxin from the prodrugs. Preliminary cell testing and reduction potential measurements support optimisation of the compounds towards tumour-selective mutual prodrugs.     

不同细胞质埃塞俄比亚芥的获得及其生物学特性比较研究

以埃塞俄比亚芥(Brassica carinata)为轮回亲本,分别对甘蓝型油菜(Brassica napus)和芥菜型油菜(Brassica juncea)连续回交7代,获得了具有核基因组基本相同而细胞质不同的新材料:N/BBCC和J/BBCC。两个新材料及其轮回亲本C/BBCC在生长发育时期、植株外部形态特征、POD酶活性等方面表现基本一致,但在雄性育性、脯氨酸含量及相对电导率上存在着明显差异,其中N/BBCC雄蕊发育不完善、花药瘦小、药内花粉量很少且多为无生活力的畸形花粉,表现严重败育;J/BBCC和N/BBCC的脯胺酸含量及相对电导率明显低于其轮回亲本C/BBCC。     

Sulfur mustard induced cytokine production and cell death: Investigating the potential roles of the p38, p53, and NF‐κB signaling pathways with RNA interference

Cutaneous and ocular injuries caused by sulfur mustard (SM; bis‐(2‐chloroethyl) sulfide) are characterized by severe inflammation and death of exposed cells. Given the known roles of p38MAPK and NF‐κB in inflammatory cytokine production, and the known roles of NF‐κB and p53 in cell fate, these pathways are of particular interest in the study of SM injury. In this study, we utilized inhibitory RNA (RNAi) targeted against p38α, the p50 subunit of NF‐κB, or p53 to characterize their role in SM‐induced inflammation and cell death in normal human epidermal keratinocytes (NHEK). Analysis of culture supernatant from 200 μM SM‐exposed cells showed that inflammatory cytokine production was inhibited by p38α RNAi but not by NF‐κB p50 RNAi. These findings further support a critical role for p38 in SM‐induced inflammatory cytokine production in NHEK and suggest that NF‐κB may not play a role in the SM‐induced inflammatory response of this cell type. Inhibition of NF‐κB by p50 RNAi did, however, partially inhibit SM‐induced cell death, suggesting a role for NF‐κB in SM‐induced apoptosis or necrosis. Interestingly, inhibition of p53 by RNAi potentiated SM‐induced cell death, suggesting that the role of p53 in SM injury, may be complex and not simply prodeath. © 2010 Wiley Periodicals, Inc. J Biochem Mol Toxicol 24:155–164, 2010; Published online inWiley InterScience ( www.interscience.wiley.com ). DOI 10.1002/jbt.20321     

茎瘤芥主要数量性状遗传力和遗传进度的初步研究

以23个茎瘤芥地方品种为试材估算了15个性状遗传力、遗传进度及遗传变异系数。结果表明各性状遗传力从大到小顺序为营养生长期、瘤茎膨大期、茎／叶、菜皮百分率、瘤茎产量、叶宽、瘤茎横径、出苗至瘤茎膨大始期、株鲜重、瘤茎纵径、叶长、菜形指数、开展度、瘤茎空心率、株高。瘤茎空心率、茎／叶、菜皮百分率和瘤茎产量的遗传变异较大,具有较大的选择潜力。在5％的选择率下,可获得较大的遗传进展。综合试验结果认为,瘤茎膨大期、营养生长期、茎／叶宜在早代进行一次性单株选择,菜皮百分率、瘤茎产量、叶宽等性状可适当加强早代选择。     

Peptide conjugates of 4-aminocyclophosphamide as prodrugs of phosphoramide mustard for selective activation by prostate-specific antigen (PSA)

In our continued effort to develop prodrugs of phosphoramide mustard, conjugates of 4-aminocyclophosphamide (4-NH₂-CPA) with three PSA-specific peptides were synthesized and evaluated as substrates of PSA. These include conjugates of cis-(2R,4R)-4-NH₂-CPA with a tetrapeptide Succinyl-Ser-Lys-Leu-Gln-OH, a hexapeptide Succinyl-His-Ser-Ser-Lys-Leu-Gln-OH, and a pentapeptide Glutaryl-Hyp-Ala-Ser-Chg-Gln-OH. These conjugates were cleaved by PSA efficiently and exclusively after the expected glutamine residue to release 4-NH₂-CPA, the activated prodrug form of phosphoramide mustard. The cleavage was most efficient for the pentapeptide conjugate 3 (Glutaryl-Hyp-Ala-Ser-Chg-Gln-NH-CPA), which showed a half-life of 55 min with PSA, followed by the hexapeptide conjugate 2 (Succinyl-His-Ser-Ser-Lys-Leu-Gln-NH-CPA) and the tertrapeptide conjugate 1 (Succinyl-Ser-Lys-Leu-Gln-NH-CPA) with half-lives of 6.5 and 12 h, respectively. These results indicate a potential of the conjugate 3 as an anticancer prodrug of phosphoramide mustard for selective PSA activation.     

叶用芥菜细胞质雄性不育相关基因orf 220的分子特性

采用榨菜细胞质雄性系为不育源,通过变种间杂交和回交的方法,转育叶用芥菜细胞质雄性不育种质。根据Brassica nap型细胞质雄性不育相关基因off222设计兼并引物,利用特异引物PCR方法,分别在不育源、F1、BC2以及BC3世代中扩增出一条600～700bp大小的特异条带,而在叶用芥菜保持系中无此条带,具细胞质基因遗传特性。进一步序列分析表明,此特异条带大小为663bp,具有起始密码子和终止密码子,编码220个氨基酸,定名为off220。同时,off220推导的氨基酸序列存在两个跨膜区,氨基端与Oenothera berteriana中的COXⅢ(线粒体细胞色素氧化酶复合体亚基)、萝卜中的ATP8(线粒体ATP酶复合体亚基)以及向日葵中的ORFB(ATP酶复合体α亚基相关)蛋白氨基端高度同源。RT-PCR方法分析表明,off220基因的表达为组成性表达,无器官特异性。