Room temperature crystal structure of the fast switching M159T mutant of the fluorescent protein dronpa

The fluorescent protein Dronpa undergoes reversible photoswitching reactions between the bright “on” and dark “off” states via photoisomerization and proton transfer reactions. We report the room temperature crystal structure of the fast switching Met159Thr mutant of Dronpa at 2.0‐Å resolution in the bright on state. Structural differences with the wild type include shifted backbone positions of strand β8 containing Thr159 as well as an altered A‐C dimer interface involving strands β7, β8, β10, and β11. The Met159Thr mutation increases the cavity volume for the p‐hydroxybenzylidene‐imidazolinone chromophore as a result of both the side chain difference and the backbone positional differences. Proteins 2015; 83:397–402. © 2014 Wiley Periodicals, Inc.     

Influence of different growth conditions on the survival and the efficacy of freeze-dried Pseudomonas fluorescens strain Pf153

High viability, storability and tolerance to variable environmental conditions are key factors in the development of microbial biological control agents (BCAs). The efficacy of microbial BCAs is influenced by the culture conditions and formulation process. Therefore, we investigated the influence of diverse growth conditions on the survival during freeze-drying and on the biocontrol efficacy of Pseudomonas fluorescens strain Pf153. Culture time, temperature and media, mild heat shock and pH change influenced the bacterium viability after freeze-drying. The best survival rate was reached by cultivation in King’s broth for 16 or 20 h. Growth temperatures of 25 and 30°C and a mild heat shock at 35°C for one hour influenced the survival rate positively. In all bioassays against Botrytis cinerea on Vicia faba leaves, Pf153 showed a significant increased efficacy compared to the untreated control. No differences of the efficacy between fresh and freeze-dried cells were observed. Furthermore, a growth temperature of 20°C increased the efficacy of Pf153 against B. cinerea. These results underline that the quality of the formulated product can be improved by manipulating the fermentation process.     

Potential effects of warmer worms and vectors on onchocerciasis transmission in West Africa

Development times of eggs, larvae and pupae of vectors of onchocerciasis (Simulium spp.) and of Onchocerca volvulus larvae within the adult females of the vectors decrease with increasing temperature. At and above 25°C, the parasite could reach its infective stage in less than 7 days when vectors could transmit after only two gonotrophic cycles. After incorporating exponential functions for vector development into a novel blackfly population model, it was predicted that fly numbers in Liberia and Ghana would peak at air temperatures of 29°C and 34°C, about 3°C and 7°C above current monthly averages, respectively; parous rates of forest flies (Liberia) would peak at 29°C and of savannah flies (Ghana) at 30°C. Small temperature increases (less than 2°C) might lead to changes in geographical distributions of different vector taxa. When the new model was linked to an existing framework for the population dynamics of onchocerciasis in humans and vectors, transmission rates and worm loads were projected to increase with temperature to at least 33°C. By contrast, analyses of field data on forest flies in Liberia and savannah flies in Ghana, in relation to regional climate change predictions, suggested, on the basis of simple regressions, that 13–41% decreases in fly numbers would be expected between the present and before 2040. Further research is needed to reconcile these conflicting conclusions.     

Gibberellic acid signaling is required for ambient temperature‐mediated induction of flowering in Arabidopsis thaliana

Distinct molecular mechanisms integrate changes in ambient temperature into the genetic pathways that govern flowering time in Arabidopsis thaliana. Temperature‐dependent eviction of the histone variant H2A.Z from nucleosomes has been suggested to facilitate the expression of FT by PIF4 at elevated ambient temperatures. Here we show that, in addition to PIF4, PIF3 and PIF5, but not PIF1 and PIF6, can promote flowering when expressed specifically in phloem companion cells (PCC), where they can induce FT and its close paralog, TSF. However, despite their strong potential to promote flowering, genetic analyses suggest that the PIF genes seem to have only a minor role in adjusting flowering in response to photoperiod or high ambient temperature. In addition, loss of PIF function only partially suppressed the early flowering phenotype and FT expression of the arp6 mutant, which is defective in H2A.Z deposition. In contrast, the chemical inhibition of gibberellic acid (GA) biosynthesis resulted in a strong attenuation of early flowering and FT expression in arp6. Furthermore, GA was able to induce flowering at low temperature (15°C) independently of FT, TSF, and the PIF genes, probably directly at the shoot apical meristem. Together, our results suggest that the timing of the floral transition in response to ambient temperature is more complex than previously thought and that GA signaling might play a crucial role in this process.     

外源NO缓解蝴蝶兰低温胁迫伤害的生理机制研究

以蝴蝶兰品种‘台湾黄金’幼苗(苗龄15个月)为试验材料,通过叶面喷施200μmol·L-1的NO供体硝普钠(SNP),低温胁迫(昼/夜:12℃/7℃)处理5d和10d,分别测定叶片电解质渗漏率、丙二醛(MDA)含量、pH、渗透调节物质含量及几种保护酶活性,探讨外源NO缓解蝴蝶兰低温胁迫伤害的生理机制。结果表明:低温胁迫条件下,预施SNP处理可以有效抑制蝴蝶兰叶片电解质渗漏率、MDA含量和pH的上升,显著提高叶片可溶性糖、可溶性蛋白及脯氨酸(Pro)含量,显著延缓超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)和抗坏血酸过氧化物酶(APX)等抗氧化酶活性的下降,增强多酚氧化酶(PPO)和苯丙氨酸解氨酶(PAL)的活性。研究认为,外源NO供体SNP可以通过保护蝴蝶兰幼苗的细胞膜系统,增加渗透调节物质含量,提高保护酶活性来减轻低温胁迫对蝴蝶兰幼苗的伤害,提高其抗低温胁迫的能力。     

茶树中2个NAC转录因子基因的克隆及温度胁迫的响应

利用茶树转录组数据库,检索得到2个NAC家族转录因子基因CsNAC1和CsNAC2。通过RT-PCR方法,将其从茶树‘迎霜’中分离克隆,利用荧光定量PCR方法,对CsNAC1和CsNAC2基因在‘迎霜’和‘安吉白茶’2个茶树品种不同组织以及温度胁迫处理下的表达进行分析,以探讨NAC家族转录因子在温度胁迫下的响应特征。结果表明:(1)CsNAC1和CsNAC2基因开放阅读框长度分别为1 044和1 047bp,分别编码347个和348个氨基酸;蛋白功能域预测和多重对比显示,CsNAC1和CsNAC2蛋白N端均含有典型NAC家族成员所具有的NAM保守结构域。(2)进化分析表明,CsNAC1和CsNAC2分别属于NAC家族的NAP和AtNAC3亚家族。(3)三维分子模型建模显示,CsNAC1和CsNAC2蛋白分别含有3个和2个α-螺旋,6个和7个β-折叠。(4)荧光定量PCR结果显示,CsNAC1在2个茶树品种中具有较相似的组织特异性,均在茶树成熟叶中表达量最高;CsNAC2则分别在‘安吉白茶’的幼叶中,‘迎霜’的根中表达量最高;高温(38℃)和低温(4℃)处理下,CsNAC1和CsNAC2基因的表达均受不同温度胁迫影响,不同茶树品种、不同时间段的表达存在差异。     

平果喀斯特山地不同植物群落的土壤质量评价

了解喀斯特山地的土壤环境质量,对其植被恢复和石漠化治理有着重要意义。基于群落土壤肥力分析,采用典范对应分析(CCA)和因子分析相结合的方法,定量评价了广西平果喀斯特山地灌丛、灌木、仪花和蚬木群落的土壤质量。结果表明:土壤物理结构随植被恢复得到改善,土壤容重下降、孔隙增多、持水能力增强。土壤p H值以灌丛最高,仪花和蚬木林的土壤有机质、全氮和碱解氮含量明显高于灌丛和灌木,灌丛、灌木的土壤全磷、全钾和速效钾含量则显著高于次生乔木林;土壤速效磷和交换性钙含量差异不显著。CCA可以有效筛选土壤质量评价指标,土壤质量综合评价分值的大小顺序为:仪花林(0.41)蚬木林(0.27)灌丛(-0.20)灌木林(-0.44)。导致灌丛和灌木林土壤质量较低的原因主要与群落生产力低、土壤有机质等养分积累少有关。     

昼夜温差对番茄生长发育、产量及果实品质的影响

在3间人工气候室精密受控环境中,保持日平均温度为22 ℃,设置昼夜温差分别为6 ℃(25 ℃/19 ℃)、8 ℃(26 ℃/18 ℃)、10 ℃(27 ℃/17 ℃),研究昼夜温差对番茄生长的影响.结果表明: 番茄不同品种、不同生长时期适宜的昼夜温差条件不同.番茄开花前,与6 ℃温差相比,8 ℃温差可显著加快野生种醋栗番茄LA1781生长发育,使幼苗株高增加23.1%,出叶加快1～2片,开花提前7 d;10 ℃温差对LA1781苗期的促进作用与8 ℃温差相似.对栽培种普通番茄LA2397和LA0490来说,6 ℃温差使幼苗生长良好,8 ℃温差对幼苗无显著促进作用;与6 ℃温差相比,10 ℃温差对苗期生长及开花有抑制作用,使株高降低12.0%～18.3%,出叶慢2～3片,开花推迟2～4 d.10 ℃温差使3个品种番茄地上部分干质量增加25.2%～44.2%.番茄开花后,与6 ℃温差相比,10 ℃温差可显著提高LA1781的产量和果实品质,使果实数增加34.7%,单株产量增加92.1%,平均单果质量增加40.0%,果实可溶性糖含量增加16.3%,番茄红素含量增加95.6%.与6 ℃温差相比,LA2397和LA0490在8 ℃温差下产量和果实品质提高,番茄红素含量增加超过2倍;在10 ℃温差下产量略有降低(5.0%),果实含糖量降低,但果实尺寸和番茄红素含量增加.表明番茄苗期生长温差不宜过大,花果期适当增大昼夜温差可提高产量和果实品质,但温差过大易造成生长不良和减产.