铁皮石斛PRC2复合体成员的鉴定及对胁迫响应的表达分析

为了探讨PRC2复合体在铁皮石斛生长发育和胁迫响应中的功能,通过生物信息学方法筛选了铁皮石斛PRC2核心成员DcCLF、DcSWN、DcEMF2、DcFIE和DcMSI1,借助酵母双杂交技术分析了它们之间的互作关系,利用半定量PCR分析了PRC2核心成员的组织表达谱,通过荧光定量PCR检测了它们对非生物胁迫(低温、高温、脱水)和病害胁迫(齐整小核菌、灰葡萄孢霉菌)的响应情况。结果表明,铁皮石斛PRC2复合体包含5个成员:E(z)同源基因DcCLF和DcSWN、Su(z)12基因DcEMF2、ESC基因DcFIE、p55基因DcMSI1,且这5个成员间的互作关系基本符合模式植物的互作模式,也存在物种特异性,表明PRC2复合体在进化中既有保守性也有特殊性。PRC2核心成员在铁皮石斛根、茎、叶、花蕾、成花中均有表达,但不同基因的表达存在组织差异性。同时,PRC2成员响应不同的环境和病害胁迫:DcCLF受低温、高温和脱水等各种环境胁迫的显著诱导;DcMSI1和DcEMF2在齐整小核菌侵染下表达明显上调,而DcSWN在灰葡萄孢霉菌侵染下受诱导程度最大。铁皮石斛PRC2复合体在生长发育和胁迫应答中发挥重要作用,可为分子辅助育种提供关键靶标基因。     

灌浆期高温胁迫对不同品种小麦蛋白组分及面团揉混特性的影响

为研究灌浆期高温胁迫对不同品种小麦蛋白组分及面团揉混特性的影响,以济麦22(JM22)和新麦26(XM26)为材料,通过灌浆初期(S1)和灌浆中期(S2)在田间搭棚进行高温胁迫处理,以未进行高温胁迫的大田小麦作对照(CK),收获后对小麦淀粉黏滞谱、蛋白质组分含量和揉混参数等进行分析。结果表明,与各自CK相比,JM22的黏滞谱参数除回复值和糊化温度降低外,其余参数均升高,XM26的黏滞谱参数除峰值时间外均降低。S1和S2使JM22的峰值黏度、低谷黏度、崩解值、最终黏度、峰值时间分别较CK提高2.81%和18.63%、7.71%和19.51%、11.88%和21.15%、1.88%和12.22%、2.45%和4.08%,且S2均大于CK和S1,S1与CK差异不显著;S1和S2使XM26的峰值黏度、低谷黏度、最终黏度、回复值分别较CK降低12.95%和31.21%、1.81%和27.18%、2.50%和22.22%、3.57%和14.39%,其中,S2、S1与CK三者之间的峰值黏度均达显著水平。与CK相比,高温胁迫后JM22的蛋白质含量降低,而XM26升高。两品种各组分蛋白含量均发生改变,S1和S2使JM22谷蛋白含量减少4.52%和6.01%,S1和S2使XM26谷蛋白增加13.66%和17.27%,不可溶蛋白增加28.95%和34.80%,谷醇比也增加。高温处理对两品种面团揉混参数值也有一定影响,S1使JM22和XM26的峰值时间、峰值高度、8 min带宽分别较各自CK显著增加8.50%和42.80%、12.20%和2.80%、26.57%和68.30%,而S2的两品种只有峰值时间同时显著增加,分别增加9.60%和28.50%。本研究结果为优质专用小麦品质提升栽培技术提供了理论基础。     

盐胁迫下宁夏枸杞Na+吸收及Na+/H+转运蛋白与H+-ATPase基因表达的研究

为从分子水平揭示宁夏枸杞钠的吸收积累机理,本试验采用原子吸收分光光度法和实时荧光定量PCR(RT-qPCR)方法,对盐胁迫下宁夏枸杞根中Na⁺、K⁺含量以及质膜和液泡膜Na⁺/H⁺转运蛋白与H⁺-ATPase基因表达水平进行测定分析。结果表明,相同胁迫时间下,随着NaCl处理浓度的增加,枸杞根系中Na⁺浓度总体呈缓慢增加趋势,K⁺含量呈先增加后减少趋势,Na⁺/K⁺比值呈先减少后增加趋势;编码质膜和液泡膜的Na⁺/H⁺转运蛋白基因LbSOS1、LbNHX1以及液泡膜H⁺-ATPase基因LbVHA-C1表达量均呈升高趋势,质膜H⁺-ATPase基因LbHA1表达量呈先升高后降低趋势。相同NaCl处理浓度下,随着胁迫时间的延长,Na⁺含量总体呈增加趋势,K⁺含量呈先增加后减少趋势,Na⁺/K⁺比值呈增加趋势。LbSOS1、LbNHX1表达量总体呈先升高后降低趋势,LbVHA-C1、LbHA1表达量总体呈降低的趋势。相关性分析显示,不同胁迫时间下,枸杞根中LbSOS1、LbNHX1、LbVHA-C1和LbHA1表达量与Na⁺含量存在一定的正相关或负相关。上述结果表明,在低浓度NaCl胁迫时,维持枸杞体内较高的K⁺/Na⁺比值是宁夏枸杞耐盐的主要方式之一,同时也说明在胁迫初期,质膜和液泡膜的Na⁺/H⁺转运蛋白与H⁺-ATPase参与了枸杞细胞中Na⁺及时排出胞外和区隔于液泡,从而保持了根细胞内Na⁺的稳定性。此外,随着胁迫时间的延长和NaCl处理浓度的增加,LbSOS1、LbNHX1、LbVHA-C1和LbHA1的表达水平均降低,而 Na⁺积累量大幅增加,致使枸杞抗盐性降低。本研究揭示了宁夏枸杞的耐盐机理,为利用宁夏枸杞改良宁夏大面积盐碱地提供了理论依据。     

碱胁迫下榆叶梅差异基因表达分析

为了解榆叶梅(Prumus triloba Lindl.)在短期碱胁迫下的分子响应机制,本研究采用Illumina HiSeq^TM2500高通量测序技术对其在0、1、3、6、12 h碱胁迫下的叶片进行了de novo转录组测序,以此鉴定与碱胁迫相关的差异表达基因(DEGs)。结果表明,从5 960万条raw reads中获得了约5 300万条高质量clean reads,经从头组装后获得了124 786条榆叶梅unigenes。基于8 948个 unigenes差异表达(DEGs), GO富集分析发现,其中有28个基因可能在早期碱胁迫响应机制中起着重要作用; KEGG富集分析发现,不同碱处理时间下的KEGG途径(pathways)具有显著差异。对7个碱性胁迫反应相关基因进行RT-qPCR验证,所得基因的表达模式与RNA-Seq数据结果一致,证实了RNA-Seq结果的可靠性。本研究首次对碱胁迫下的榆叶梅叶片进行了转录组分析,获得的序列数据极大地丰富了榆叶梅可利用的基因资源,深入了解了榆叶梅碱胁迫的分子响应机制,同时为研究以榆叶梅为砧木的中国李(Prunus salicina)在碱胁迫下的适应机制奠定了分子研究基础。     

谷子MDH基因非生物逆境响应特性研究

为揭示谷子MDH基因对逆境胁迫的响应,本研究通过序列比对得到2个谷子苹果酸脱氢酶(malate dehydrogenase, MDH)基因SiMDH1、SiMDH2,并对SiMDH1和SiMDH2的编码蛋白特征、基因结构、功能、进化等性状进行了系统的分析和预测,用荧光实时定量PCR(qPCR)检测了SiMDH1和SiMDH2在苗期不同逆境及关键生育期干旱胁迫和不同光照强度下的表达。蛋白特征参数和序列分析表明,SiMDH1和SiMDH2蛋白特征参数相似度很高,序列非常保守,都含有MDH基因的特征功能域苹果酸酶NAD结合域和苹果酸酶C端功能域。亚细胞定位预测显示,SiMDH1和SiMDH2主要定位于叶绿体、细胞质和线粒体。启动子区域顺式元件分析发现,在SiMDH1和SiMDH2启动子区域含有大量光应答、激素类和其他类应答元件。苗期逆境qPCR表达谱分析发现,SiMDH1和SiMDH2受脱落酸(ABA)、聚乙二醇(PEG)、高盐和低温不同程度诱导,揭示SiMDH1和SiMDH2参与了谷子苗期非生物逆境胁迫应答。进一步分析发现,SiMDH1和SiMDH2参与了谷子在拔节、抽穗、灌浆期的干旱应答,而光照强度会显著影响SiMDH1和SiMDH2基因在不同生育期的表达。本研究为进一步分析MDH逆境应答机制以及利用基因工程方法改善作物抗逆性提供了试验和理论支持。     

基于粒子图像测速的坡面流水动力学特性

粒子图像测速(Particle Image Velocimetry, PIV)技术具有多点同时测量、对水流无干扰的优点,该研究利用高分辨率PIV(分辨率为64 pixels/mm),测量了7组坡面流(水深范围为0.5～1.1 cm,雷诺数范围为1 000～3 000),并测量1组深水明渠紊流作为对照,研究了流速轮廓线和修正系数、紊动强度和雷诺应力、偏态系数和峰度系数的变化规律。结果表明:1)PIV能够有效观测坡面流床面至水面的流速分布。当坡面流流态为过渡流时,流速修正系数随着雷诺数的增加呈对数增加,均值为0.77;2)对比深水明渠紊流的紊动强度,坡面流的流向紊动强度较大,而垂向紊动强度较小,且随着水深及雷诺数的增加,坡面流紊动强度逐渐与深水明渠紊流的特征吻合。深水明渠紊流中受雷诺应力影响的流体占比约80%,而坡面流中受雷诺应力影响的流体占比小于80%,随着雷诺数的增加坡面流中受雷诺应力影响的流体占比变大;3)对比深水明渠紊流的峰度系数,坡面流的峰度系数大部分大于3,表明坡面流较深水明渠紊流出现极端流速事件的概率小。PIV技术有利于实验室研究坡面水力侵蚀的力学机理机制问题。     

2种紫花苜蓿耐盐生理特性的初步研究

为研究2种紫花苜蓿(Medicagosativa L.)“阿迪娜”(耐盐基因型)和“秘鲁”(敏盐基因型)在盐胁迫处理下的生理特性与耐盐机理,采用150mmol/LNaCl胁迫处理2种紫花苜蓿幼苗,分别测定盐处理前和处理后2,4,6,8,16h的过氧化氢酶(CAT)、过氧化物酶(POD)、丙二醛(MDA)、脯氨酸(PRO)、相对含水量(RWC)和叶绿素(Chl)含量。结果表明:2种紫花苜蓿的过氧化氢酶活性、过氧化物酶活性和丙二醛含量在盐胁迫下总体呈上升趋势,脯氨酸含量和相对含水量表现出相反的趋势。叶绿素含量在盐胁迫前期稳定不变,但在胁迫后期显著性下降;2种紫花苜蓿的相对含水量和叶绿素含量的变化趋势与植株表型变化相对应。主成分分析表明,过氧化氢酶和丙二醛的贡献率最大,能更好地为苜蓿耐盐机理及分子育种研究提供理论依据。     

干旱胁迫下腐植酸对燕麦叶绿素荧光特性的调控效应

【目的】明确腐植酸(HA)对干旱胁迫下燕麦叶片叶绿素荧光特性的调控效应。【方法】采用盆栽试验,研究了在正常供水(75%田间持水率)、中度干旱胁迫(60%田间持水率)、重度干旱胁迫(45%田间持水率)3个水分条件下喷施HA对燕麦叶片叶绿素量及荧光参数的影响。【结果】①水分胁迫导致Chla+Chlb、Chla/Chlb、Fm、Fv、Fv/Fm、Fv/Fo和qP显著降低,而Fo和NPQ显著升高;②与CK相比,干旱胁迫下HA处理Chla+Chlb提高0.6%~40.82%、Chla/Chlb提高1.13%~30.09%、Fm提高0.7%~121.19%、Fv提高1.0%~171.79%、Fv/Fm提高0.2%~83.89%、Fv/Fo提高1.9%~211.56%、qP提高0.1%~68.30%、NPQ提高6.02%~73.36%、而Fo降低0.70%~14.06%,其中在重度干旱胁迫下均达到显著差异。【结论】干旱胁迫对燕麦PSⅡ光反应系统产生明显伤害,喷施腐植酸可缓解其影响,且在重度干旱胁迫条件下效果最明显。     

Transpiration efficiency of Amaranth (Amaranthus sp.) in response to drought stress

Drought is a major abiotic stress responsible for severe crop losses worldwide. Development of new crop varieties with increased drought tolerance is one way to increase crop productivity. The aim of this study was to characterise the diversity of nine accessions belonging to Amaranthus tricolor and A. cruentus, in response to drought stress using a dry-down protocol to characterise the transpiration efficiency (TE). Plants were subjected to either a gradual dry down or well-watered conditions. Results showed that TE was significantly higher (P < 0.01) in water-deficient (WD) plants compared to water-sufficient (WS) plants, 2.40 g kg^?1–7.13 g kg^?1 and 2.19 g kg^?1–4.84 g kg^?1, respectively. There was no significant difference in the fraction of transpirable soil water (FTSW) threshold decline between the amaranth genotypes. TE was highly correlated with yield under both WS (r = 0.89, P < 0.001) and WD conditions (r = 0.662, P < 0.001), and negatively correlated with root-to-shoot ratio under both WS (r = ?0.488, P < 0.05) and WD conditions (r = ?0.460, P < 0.05). Significant genotypic differences were seen for growth rate and stress susceptibility index (SSI). The result obtained in this investigation underline the need to identify genotypic variation in water use efficiency in amaranth.

Abbreviations: FTSW: Fraction of transpirable soil water; NTR: normalised transpiration rate; SSI: Stress susceptibility index; TE: transpiration efficiency; WHC: water holding capacity; WD: water-deficient; WS: water-sufficient.     

Trx-1 over-expression attenuates oxidative stress injury in MPP+ -induced PC12 cells by regulating NF-κB signaling pathway

AIM:To investigate the inhibitory effect of thioredoxin 1 (Trx-1) over-expression on oxidative stress injury in 1-methyl-4-phenylpyridinium (MPP⁺)-induced rat pheochromocytoma PC12 cells by regulating NF-κB signaling pathway.METHODS:The PC12 cells were damaged by treatment with MPP⁺ at 1, 3 and 5 mmol/L, and the optimal concentration of 3 mmol/L was selected. The cell viability was measured by MTT assay. The oxidative stress indexes lactate dehydrogenase (LDH) activity, superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in the cell culture supernatant were detected, and the protein expression of Trx-1 was determined by Western blot. Lentiviral infection with Ad-Trx-1-GFP sequence was used to establish a model of MPP⁺-treated PC12 cells with Trx-1 over-expression. The effects of Trx-1 over-expression on the cell viability, oxidative stress responses and NF-κB signaling pathway were determined by MTT assay, commercial kits and Western blot. The effects of phorbol 12-myristate 13-acetate (PMA), an activator of NF-κB signaling pathway, on the viability and oxidative stress of PC12 cells were observed. The NF-κB signaling pathway inhibitor pyrrolidine dithiocarbamate (PDTC) was used in MPP⁺-treated PC12 cells with Trx-1 over-expression, and the cell viability and oxidative stress responses were measured. RESULTS:The viability of PC12 cells, SOD activity in the supernatant and protein expression of Trx-1 were decreased, while LDH activity and MDA content in the supernatant were increased significantly by treatment with MPP⁺ at 1, 3 and 5 mmol/L. The effect of MPP⁺ at 3 mmol/L and 5 mmol/L was significantly greater than that at 1 mmol/L (P<0.05), and no significant difference between 3 mmol/L and 5 mmol/L was observed (P>0.05). The inhibitory effect of MPP⁺ on the viability of PC12 cells, and the oxidative stress injury and activation of NF-κB signaling pathway induced by MPP⁺ were significantly attenuated by over-expression of Trx-1. The inhibitory effect of MPP⁺ on the viability of PC12 cells and the oxidative stress injury induced by MPP⁺ were promoted by the activation of NF-κB signaling pathway, while the protective effects of Trx-1 over-expression on the MPP⁺-treated PC12 cells were enhanced by the inhibition of NF-κB signaling pathway. CONCLUSION:Over-expression of Trx-1 protects MPP⁺-treated PC12 cells from oxidative stress injury by regulating NF-κB signaling pathway.