Plasmid composition and the chpG gene determine the virulence level of Clavibacter capsici natural isolates in pepper

The gram-positive bacterial species Clavibacter capsici causes necrosis and canker in pepper plants. Genomic and functional analyses of C. capsici type strain PF008 have shown that multiple virulence genes exist in its two plasmids. We aimed to identify the key determinants that control the virulence of C. capsici. Pepper leaves inoculated with 54 natural isolates exhibited significant variation in the necrosis. Six isolates showed very low virulence, but their population titres in plants were not significantly different from those of the highly virulent isolates. All six isolates lacked the pCM1_Cc plasmid that carries chpG, which has been shown to be required for virulence and encodes a putative serine protease, but two of them, isolates 1,106 and 1,207, had the intact chpG elsewhere in the genome. Genomic analysis of these two isolates revealed that chpG was located in the pCM2_Cc plasmid, and two highly homologous regions were present next to the chpG locus. The chpG expression in isolate 1,106 was not induced in plants. Introduction of chpG of the PF008 strain into the six low-virulence isolates restored their virulence to that of PF008. Our findings indicate that there are at least three different variant groups of C. capsici and that the plasmid composition and the chpG gene are critical for determining the virulence level. Moreover, our findings also indicate that the virulence level of C. capsici does not directly correlate with bacterial titres in plants.     

The histology of mature gonads of Oreochromis (Nyasalapia) karongae (Trewavas)

Histology of gonads of Oreochromis karongae was undertaken to study internal cell characteristics during maturation. This study was necessitated by low spawning output of the fish species. Several oocyte stages, ranging from primary forms to vitellogenesis, suggest that the maturation was generally succesfully attained in the fish ponds. Pre‐vitellogenesis oocytes (oogonia to perinuclear stage) and more advanced vitellogenesis (primary vesicle to tertiary yolk vesicle) oocyte stages were all found in the same gonads. However, there were some discontinuities observed during stages 3 and 4, suggesting selective maturation. Failure of gonads to mature normally is attributed to an ecological crunch that was in a previous study associated with environmental factors. Atretic oocytes were also recorded in the same gonads, a sign that some oocytes failed to mature normally. This indicates insufficient stimuli for normal gonad development. Several stages of spermatogenesis (spermatocytes, spermatid and spermatozoa) were also found in the same gonads. Selective recrudescence was more pronounced in O. karongae because generally less oocytes attained final maturation stages compared to Oreochromis niloticus and other tilapias. This could be the main reason for low natural breeding that has been observed in both wild and captive stocks, and led to the abandonment of its use in aquaculture. This study corroborates findings of previous studies that depended solely on external gonad characteristics. Histology provides conclusive evidence from internal cell characteristics that other techniques are unable to show.     

昆明西山滇青冈林内滇青冈种子库动态的研究

通过对昆明西山滇青冈林内滇青冈种子库的跟踪取样调查和种子埋藏试验,对滇青冈种子库的动态进行了研究。昆虫在种子成熟前侵入种子,经种子雨进入种子库时已有71.8%的种子失去萌发能力。种子雨输入种子库的绝大部分种子停留在表面种子库,其中48.55%的种子被虫害,25.36%被某些非生物或生物搬运,17.39%的腐烂,8.7%的被动物当场取食,没有种子萌发,影响种子库动态的各种因子的作用大小在时间上是变化。被搬运的种子中,有4.9%的由表面种子库转移到埋藏种子库。土层是滇青冈种子的安全生境,土壤种子库的存在时间超过250天。埋入土壤的试验种子一直处于静止状态,到6月雨季后有80%种子萌发,20%的腐烂。萌发种子数是当年产种子的0.26%。滇青冈林内的滇青冈种子库是季节性的,种子库对种群个体的补充作用是有限的。     

电穿孔介导质粒DNA肿瘤内转移抑制恶性肿瘤生长与转移

利用携带绿色荧光蛋白（green fluorescent protein, GFP）编码基因的表达质粒,测试电穿孔方法介导目的基因活体组织内转移的效率并优化电击参数.在此基础上采用电穿孔技术直接将编码白介素12（IL-12）、白介素2（IL-2）、粒单细胞克隆刺激因子（GM-CSF）等免疫调节因子或反义血管内皮细胞生长因子121（VEGF₁₂₁）、可溶性血管内皮细胞膜受体（sFlk-1及ExTek）等血管生成抑制因子表达质粒转移至肿瘤局部.实验结果表明电穿孔介导GFP表达质粒肌肉内转移的效率较高,GFP可在肌细胞内持续高水平表达3周以上,而在肿瘤细胞内只能表达4～6 d,但高电压短脉冲电击组肿瘤内GFP阳性细胞数比低电压长脉冲组高2.68倍.多次电击介导IL-12表达质粒转移至肿瘤组织内,可有效地抑制小鼠膀胱癌BTT-gfp、人乳腺癌MCF-7及肝癌SMMC 7721-gfp的生长.MCF-7对血管生成抑制因子基因转移治疗较敏感,单独应用反义VEGF₁₂₁、sFlk-1或ExTek即显示明确的治疗效果.SMMC 7721-gfp单独应用sFlk-1有效.小鼠膀胱癌对单独应用反义VEGF₁₂₁、sFlk-1或ExTek治疗效果不理想,但联合应用sFlk-1和ExTek仍然可以有效地抑制肿瘤生长与转移,甚至使肿瘤缩小或消失.提示电穿孔技术是一项高效、安全、经济的体内基因转移方法.     

Detection of hatching inhibitors and hatching factor stimulants for golden potato cyst nematode, Globodera rostochiensis, in potato root leachate

In a comparison of four potato varieties, in-soil hatch of the golden potato cyst nematode (Globodera rostochiensis) was positively correlated to in vitro hatch in response to potato root leachate (PRL). The in-soil hatch of cysts of G. rostochiensis to two of the four varieties was significantly less than that of the control (cysts in gravel without potato plants) in the first 2 wk after plant emergence, suggesting the production of hatching inhibitors (HIs) by young potato plants. The hatching factor: hatching inhibitor ratio of PRL was positively associated with the net hatching activity of the PRL. Four zones of HI activity were resolved following gel permeation chromatography of PRL on Sephadex G-10. Hatch-inactive chemicals, which stimulated the activity of hatching factors (HFs) in PRL (hatching factor stimulants, HSs), were also isolated from PRL, hatch levels induced by individual HFs responding differently to the same HS preparation. The complex interactions between individual HFs and other hatching chemicals in PRL was illustrated when addition of the hatch-active potato glycoalkaloid α-solanine caused both inhibition and stimulation of PRL-induced hatch, depending on the α-solanine concentration.     

环境因素所致印迹基因改变与子代器官发育

印迹基因是由大约100个基因组成的一类特殊子集,主要以亲本单等位基因的方式表达,对胚胎的生长发育具有重要作用.近年来发现,环境因素所引起的印迹基因表观遗传修饰改变可造成胎儿多脏器发育不良甚至成年后多疾病易感,且存在多代遗传效应.本文基于国内外最新研究进展,总结了印迹基因表达改变对个体发育阶段以及生命后期器官功能的影响,...     

南北样带温带区栎属树种种子化学组成与气候因子的关系

选取南北样带温带区8个核心分布区的7种优势栎属树种作为对象,通过检测种子的化学组成,包括淀粉、蛋白质、油脂、可溶性总糖、单宁和脯氨酸,分析其化学组成与气候因子之间的关系,结果发现:在气候因子中,温度是影响种子化学组成的主要因素,日照时数次之,而降水没有明显影响.在南北样带温带区,随纬度的增加,温度降低,年日照时数增加,生长积温减少,生长季缩短,淀粉含量积累下降,并趋向于水解为可溶性糖类;同时,从南到北,为适应低温的环境条件,栎属树种通过增加种子蛋白质、可溶性总糖和脯氨酸的含量,以提高种子的抗寒性.栎属树种种子的化学组成之间存在一定的相互关系,可溶性总糖、蛋白质和脯氨酸之间呈正相关关系,它们三者与淀粉均为负相关,种子化学组成之间的这种相互关系是植物对环境适应的生态策略.     

岩溶区青冈栎的年轮特征及其与环境因子的关系

为揭示岩溶区植物生长特征及其与环境因子的关系,利用年轮分析方法研究了岩溶区广泛分布的乔木树种青冈栎的年轮特征及其与环境因子的关系.结果表明:青冈栎年轮宽度和年轮指数的平均值分别为(2.565±0.028) mm和1.108±0.012.随着胸径的增大,年轮宽度和年轮指数在逐渐增大,年轮指数与气候因子的变化有着相关关系,与前一年6月份、12月份以及当年12月份的平均温度呈显著正相关,而与前一年4月份、10月份以及当年4月份的降雨量和前一年8月份、9月份以及当年2月份的日照总时数呈显著负相关.表明岩溶区青冈栎的生长可能主要受到温度变化的影响,降雨量和日照时数的变化也影响着青冈栎的年轮特征.温度,降雨量和日照时数等环境因子共同制约着岩溶区青冈栎的生长.     

Harnessing mesenchymal stem cell secretome: Effect of extracellular matrices on proangiogenic signaling

The low engraftment and retention rate of mesenchymal stem cells (MSCs) at the target site indicates that the potential benefits of MSC-based therapies can be attributed to their paracrine signaling. In this study, the extracellular matrices (ECMs) deposited by bone marrow-derived human MSCs in the presence and absence of ascorbic acid was characterized. MSCs were seeded on top of decellularized ECM (dECM) and the concentrations of proangiogenic and antiangiogenic molecules released in culture (conditioned) media was compared. Effects of ECM derived from MSCs with different passage numbers on MSC secretome was also investigated. Our study revealed that the expression of proangiogenesis-related factors were upregulated when MSCs were harvested on dECMs, irrespective of media supplementation, as compared with those cultured on tissue culture plates. In addition, dECM generated in the presence of ascorbic acid promoted the expression of proangiogenic molecules as compared with dECM-derived in absence of media supplementation. Further, it was observed that the effectiveness of dECM to stimulate proangiogenic signaling of MSCs was reduced as cell passage number was increased from P3 to P5. The proliferation as well as capillary morphogenesis of human umbilical vein endothelial cells (HUVECs) in the presence of conditioned media were enhanced compared with the normal HUVECs culture media. These data indicate that the secretory signatures of MSCs and consequently, the therapeutic efficacy of MSCs can be regulated by presentation of dECM composition and variation of its composition.