Physiological metabolism of Haliotis discus hannai Ino under different light qualities and cycles

The goal of this experiment was to examine the effects of different light qualities (red, white, blue) and cycles (12L:12D, 8L:16D, 4L:20D, 0L:24D, 16L:8D) on the survival, growth, metabolism and antioxidant defence system of Haliotis discus hannai Ino. Under red and white light at 4L:20D, the body weight‐specific growth rate of abalones was significantly higher than that of the 0L:24D group (P < 0.05). At 16L:8D, succinate dehydrogenase activity decreased and lactate dehydrogenase and alanine aminotransferase activity and the lactic acid content significantly rose (P < 0.05) in the red and white light groups. Under blue light, reactive oxygen species and malondialdehyde content for each light cycle were significantly higher than those of the red and white groups (P < 0.05). Under blue, white and red light, total antioxidant capacity (T‐AOC), superoxide dismutase (SOD) activity, glutathione peroxidase (GP_X) activity and reduced glutathione (GSH) content gradually rose with increased duration of light exposure. Under red light, significant differences in these parameters were detected between the 4L:20D and 16L:8D groups (P < 0.05). Under blue and white light at 16L:8D, values of T‐AOC, SOD, GP_X activity and GSH content were significantly lower than those of the 12L:12D group (P < 0.05). Overall, these results indicate that red light at light cycles of 4L:20D and 8L:16D is optimal for the culture and production of H. discus hannai, as these conditions promoted food intake, growth and health maintenance as well as normal physiological metabolism of the organism.     

茄子反向温敏雄性不育系可溶性糖含量及相关基因表达分析

为揭示茄子反向温敏核雄性不育(rTGMS)发生与糖类代谢的关系,以茄子rTGMS系‘05ms’及温度不敏感系‘S63’为试材,对不育期和可育期的叶片及花蕾的可溶性糖含量进行比较,结果表明,‘05ms’在不育和可育时期叶片和花蕾中的可溶性糖含量均显著低于‘S63’;进一步对‘05ms’花蕾进行转录组测序及对其糖类代谢相关途径进行分析,发现差异表达基因主要富集在3个通路:淀粉和蔗糖代谢、糖酵解/糖异生和氨基糖和核苷酸糖代谢,其中淀粉和蔗糖代谢通路的差异基因最多且大部分在低温条件表达下调,主要影响了葡萄糖和半乳糖醛酸的合成。此结果初步揭示了茄子反向温敏雄性不育系中糖类代谢响应温度变化的特点,为茄子雄性不育杂交育种提供理论依据。     

玉米pTAC2影响苗期叶片叶绿素合成的转录组分析

【目的】叶绿素是参与光合途径最为重要的光合色素。叶绿体的发育及叶绿素的合成在很大程度上依赖于质体基因组与核基因组之间的双向信号传导来精确协调基因表达。通过对白化表型的CRISPR/Cas9-ZmpTAC2转基因阳性纯合突变材料进行RNA-seq研究,筛选和鉴定参与叶绿素合成的相关基因,为明确叶绿素的合成途径奠定基础。【方法】以CRISPR/Cas9-ZmpTAC2玉米转基因编辑纯合突变株系为研究材料,使用透射电镜观察叶绿体超微结构和分光光度法测定叶片叶绿素含量,确定叶绿体发育状态及叶绿素合成情况。对转基因阴性材料（CK）和CRISPR/Cas9-ZmpTAC2转基因纯合编辑材料（zmptac2）苗期叶片取样进行转录组测序。通过生物信息学分析,寻找CK与zmptac2间差异表达的基因;qRT-PCR对差异表达基因进行验证。通过酵母双杂交筛选与玉米pTAC2互作的蛋白质。【结果】共获得15株T₀转基因植株,包括绿色植株（7株）和白色植株（8株）。绿色幼苗中3株为转基因阴性材料,4株为转基因阳性（2株为未编辑,2株为杂合编辑突变）,白色植株(8株)均为转基因阳性纯合编辑。与CK相比,突变体（zmptac2）叶绿体发异常,叶绿素含量显著降低。RNA-seq的结果显示,CK与zmptac2之间共检测到1 367个基因差异表达,其中618个基因上调表达（zmptac2/CK）,749个基因下调表达（zmptac2/CK）。GO富集分析显示,下调基因显著富集到叶绿体和质体中。KEGG分析表明下调表达基因显著富集在苯丙氨酸代谢、酪氨酸代谢和异喹啉生物碱生物合成等途径。选取的15个差异基因表达模式均与测序数据相一致,表明测序结果是可靠的。与CK相比,zmptac2中依赖PEP（plastid-encoded RNA polymerase）转录的基因表达量显著降低,而依赖NEP（nuclear gene-encoded RNA polymerase）转录的基因表达量则显著上升。通过对玉米cDNA文库筛选和互作验证,鉴定出ZmpTAC3与ZmpTAC2存在互作。【结论】ZmpTAC2突变会导致叶绿体早期生物合成受阻,该基因参与叶绿体发育及叶绿素合成,且该种作用是由ZmpTAC2调控PEP相关基因表达而实现的。     

两个褐飞虱海藻糖转运蛋白基因的结构及调控海藻糖代谢功能

【目的】海藻糖是昆虫中的主要血糖物质,在昆虫的发育及生理活动中发挥重要功能。其中,海藻糖转运蛋白（Tret）在将海藻糖从其生成组织（例如脂肪体）运输到其消耗组织的过程中起着重要作用。本研究通过分析褐飞虱（Nilaparvata lugens）两条Tret1的序列结构,进一步抑制NlTret1的表达,探讨这两个NlTret1在褐飞虱体内的生物学功能。【方法】以褐飞虱两条Tret1序列为研究对象,利用生物信息学技术分析其蛋白结构以及与其他昆虫之间的同源性。采用RNAi（RNA interference）技术将合成的外源dsRNA（double-stranded RNA）注射到实验室饲养褐飞虱种群体内,抑制其体内NlTret1的表达,分别在注射48 h后取材,抽取总RNA并用反转录试剂盒合成第一链cDNA,采用qRT-PCR（quantitative real-time PCR）技术检测dsNlTret1的干扰效果以及RNAi后褐飞虱体内海藻糖代谢通路中相关基因的表达,最后测定葡萄糖、海藻糖和糖原含量以及海藻糖酶活性。【结果】生物信息学分析表明,NlTret1-like X1和NlTret1-2 X1的开放阅读框长度分别为1 920和1 578 bp,分别编码639和525个氨基酸,预测蛋白分子量分别为69.29和58.71 kD,等电点分别为8.32和8.36;NlTret1-like X1和NlTret1-2 X1的二级结构主要包含螺旋和卷曲;保守结构域分析显示它们均属于MFS家族。进化分析结果显示,不同昆虫的Tret1蛋白具有较高的同源性,且褐飞虱与其他半翅目昆虫具有较近的亲缘关系;与注射dsGFP组相比,注射靶标基因的dsRNA后均能够显著沉默本基因的表达;褐飞虱体内糖原、葡萄糖在注射dsNlTret1-like X1和dsNlTret1-2 X1后,其含量均无显著变化,然而不同于注射dsNlTret1-2 X1组,在注射dsNlTret1-like X1后褐飞虱体内海藻糖含量极显著升高;干扰NlTret1-like X1 48 h后褐飞虱体内TPS1、TPS2、TRE1-1、TRE1-2和TRE2的表达均极显著下调;而干扰NlTret1-2 X1 48 h后褐飞虱体内TPS1、TPS2和TRE1-1的表达虽也极显著下降,但TRE1-2和TRE2极显著上调;在注射dsNlTret1-like X1后,试虫可溶性海藻糖酶和膜结合型海藻糖酶的活性均极显著降低,而在注射dsNlTret1-2 X1后无显著变化。【结论】褐飞虱的两个Tret1在不同组织间发挥着不同的功能,其中NlTret1-like X1在特异性转运海藻糖参与能量供应中起到更为显著的作用。研究结果有助于探索Tret1在昆虫或无脊椎动物中调控海藻糖代谢平衡的调节机制,可为将来通过调控血糖平衡来控制褐飞虱等害虫提供理论依据。     

Flow alteration and thermal pollution depress modelled growth rates of an iconic riverine fish,the Murray cod Maccullochella peelii

The serial discontinuity concept (SDC) proposes that hypolimnetic‐releasing impoundments cause major disruptions to the naturally occurring physical, chemical and biological gradients of rivers but that this impact diminishes with distance downstream. Such a gradient in discharge, flow velocity and temperature regime occurs below a large hypolimnetic‐releasing impoundment, the Hume Dam, on the River Murray in south‐eastern Australia. To examine the effects of this disturbance gradient on a warm‐water large‐bodied freshwater fish, the Murray cod (Maccullochella peelii), a bioenergetics model was developed and calibrated to explore energy expended under differing water velocities and temperature regimes. Model simulations predicted negative growth of juveniles directly downstream of the impoundment, due largely to the energetic costs associated with active and, to a lesser extent, standard metabolism outweighing the achievable energetic gains through food consumption. As flow velocity and temperature regimes became more favourable downstream, so did the simulated growth of the species. It was not until +239 km downstream of the impoundment that the model predicted that flow velocity and temperature regimes were suitable for greater weight gains. The modelled growth responses of juvenile Murray cod are consistent with the predictions of the SDC, emphasising that changes in the bioenergetics of individuals are likely to be reflected in reduced growth rates under the changed flow velocity and temperature regimes imposed by disturbance gradients. This research represents a valuable step in the biological understanding of Murray cod within variable riverine environments and emphasises the urgency required to mitigate impacts associated with hypolimnetic impoundments.     

水肥调控对滴灌马铃薯生长、品质及水肥利用的影响

研究生育期土壤水分下限调控和施肥对陕北榆林地区滴灌马铃薯生长、品质和水肥利用的影响,以期探寻马铃薯优质高产的水肥调控模式。以马铃薯紫花白为材料,分别在苗期、块茎形成期、块茎膨大期、淀粉积累期、成熟期设置3个土壤水分调控下限水平:W1(55%、60%、65%、55%、55%)、W2(65%、70%、75%、65%、65%)、W3(75%、80%、85%、75%、75%),4个施肥水平N-P_2O_5-K_2O (kg·hm~(-2))分别为F1(100-40-150 kg·hm~(-2))、F2(150-60-225 kg·hm~(-2))、F3(200-80-300 kg·hm~(-2))、F4(250-100-375 kg·hm~(-2)),共12个处理,在生育期内对马铃薯生长、产量及品质等指标进行观测,分析马铃薯各指标对水肥的响应机制。结果表明:(1)在同一施肥水平下,W2灌水处理的马铃薯生长、产量、品质、水肥利用效率(WUE)均显著高于W1和W3处理。W2处理的平均产量为43 187.15 kg·hm~(-2),比W1和W3分别增加了24.59%和5.26%,淀粉和维生素C含量分别增加了15.32%和6.37%,8.04%和4.66%,WUE分别增加了15.53%和7.54%,肥料偏生产力(PFP)分别增加了21.65%和4.41%。(2)在同一灌水水平下,F3施肥处理的马铃薯生长、产量、WUE均显著高于F1、F2和F4处理,淀粉含量和维生素C含量在F2处理最高。F3处理的平均产量为44 691.32 kg·hm~(-2),比F1、F2和F4处理分别增加了41.79%、10.98%和6.34%。WUE随施肥量的增加均呈抛物线变化趋势,PFP随施肥量增加呈减小趋势,F1处理的PFP平均为108.69 kg·kg~(-1),比F2、F3和F4处理分别显著增加了17.41%、41.06%和87.50%。(3)运用主成分分析法评价马铃薯品质表明,F3W2处理排名第一,是榆林地区马铃薯优质高产的最优水肥组合。     

烯效唑浸种对干旱胁迫下工业大麻幼苗形态、渗透调节物质及内源激素的影响

选用0.4 mg·L~(-1)的烯效唑(S_(3307))浸种工业大麻"火麻一号"种子,采用盆栽试验方法,于大麻三叶期设置清水浸种后正常供水(CK)、清水浸种后干旱胁迫(D)和烯效唑浸种后干旱胁迫(D+S)3个处理,探讨烯效唑浸种对干旱胁迫下工业大麻幼苗形态、渗透调节物质及内源激素的影响。结果表明:与D处理的植株相比,D+S处理显著提高了根系干、鲜重,分别提高46.67%～61.54%和16.46%～25.53%;恢复了胁迫后期的地上部干、鲜重,复水4 d后地上部干、鲜重较D处理分别提高了4.38%和2.23%;促进了根系生长的能力,干旱胁迫8 d后,D+S处理较D处理根长、根表面积、根体积、根总投影面积、分枝数、交叉数和根尖数分别增加了34.48%、34.77%、69.10%、70.00%、29.62%、54.28%和33.07%;提高了幼苗叶片SPAD值,降低了细胞膜透性,增加了可溶性糖和可溶性蛋白含量,干旱胁迫8 d后D+S处理较D处理SPAD值增加了28.30%,细胞膜透性减少了17.22%,可溶性糖和可溶性蛋白含量分别增加了17.32%～36.78%和5.07%～7.94%。干旱胁迫8 d后,D+S处理使工业大麻叶片中脱落酸(ABA)含量增加了1.02倍,水杨酸(SA)和茉莉酸(JA)含量分别降低了17.79%和14.40%。可见,烯效唑浸种能通过调控工业大麻幼苗生长及生理指标来增强其抗旱能力,缓解干旱胁迫对幼苗造成的伤害。