Generation and characterization of aptamers against grass carp reovirus infection for the development of rapid detection assay

Grass carp reovirus (GCRV) causes devastating viral haemorrhagic disease in farmed grass carp (Ctenopharyngon idellus). As novel molecular probes, aptamers have been widely applied in rapid diagnosis and efficient therapies against virus or diseases. In this study, three single‐stranded DNA (ssDNA) aptamers were selected against GCRV‐infected CIK cells via SELEX (systematic evolution of ligands by exponential enrichment technology). Secondary structures predicted by MFOLD indicated that aptamers formed stem‐loop structures, and GVI‐11 had the lowest ΔG value of ?30.84 KJ/mol. Three aptamers could specifically recognize GCRV‐infected CIK cells, with calculated dissociation constants (Kd) of 220.86, 176.63 and 278.66 nM for aptamers GVI‐1, GVI‐7 and GVI‐11, respectively, which indicated that they could serve as specific delivery system for antiviral therapies. The targets of aptamers GVI‐1, GVI‐7 and GVI‐11 on the surface of GCRV‐infected cells could be membrane proteins, which were trypsin‐sensitive. Furthermore, FAM‐labelled aptamer GVI‐7 could be applied to detect GCRV infection in vivo. It is the first time to generate and characterize aptamers against GCRV‐infected cells. These aptamers have great potentials in development of rapid diagnosis technology and antiviral agents against GCRV infection in aquaculture.     

混播比例和施氮肥对箭筈豌豆/燕麦草地根系特性的影响

豆科和禾本科牧草混播是栽培草地的重要模式之一,能显著改善土壤,提升系统可持续生产能力。牧草根系是土壤有机质返还的重要来源,但混播草地中牧草根系特性的变化尚不清楚。以箭筈豌豆和燕麦草地为对象,研究了不同施氮肥水平和混播比例下其根生物量、根性状的变化。结果表明:1)混播比例和施氮肥对草地根生物量和根性状有显著交互作用,混播草地根生物量显著大于单播;与高氮肥处理相比,根生物量在低、中氮肥处理下更高;高氮肥对根长、比根长、根表面积和比根面积在植物生长前期表现为抑制作用,在生长后期随混播中箭筈豌豆比例的增大,抑制作用减轻。2)混播条件下土壤硝态氮含量大于单播,而土壤铵态氮含量仅在乳熟期时大于单播,且随混播中箭筈豌豆比例增大,矿质氮含量增加;施氮肥对0~30 cm土层中的矿质氮含量有显著影响,其含量在施氮肥100 kg·hm^-2下最高,不施肥下最小。3)根生物量与土壤硝态氮含量间显著正相关,根长、根表面积、根体积与土壤硝态氮含量间显著负相关。综上,混播中箭筈豌豆比例的增加有助于改善土壤矿质氮,优化混播草地根性状。     

严格意义的无霜期统计方法探讨

根据霜冻和无霜期对农作物生长的影响及农业气候区划的指导意义,比较无霜期与严格意义上的“无冻期”的关系和区别,分析霜冻出现的初、终日与霜、结冰现象及气温、地面温度、草面温度≤0℃出现的初日、终日之间的关系,探讨“无霜冻期”的合理统计方法,并通过对鄂东地区的麻城、浠水、黄石三地的无霜期和“无冻期”的统计,总结两者的差异和特征,从而为正确理解和统计无霜冻期,提出一套新的观点和方法,为指导农业生产和农业气候区划提供依据。     

Assessing the efficacy and impact of management of an invasive species in a protected area: Poa annua on sub‐Antarctic Macquarie Island

Plant eradication is difficult, particularly in remote, protected areas. The Southern Ocean Islands are very isolated and highly protected, but the flora contains many alien plants. Small restricted populations have been eradicated, but eradication of established species has proven difficult. A better understanding of the efficacy of control methods at sub‐Antarctic temperatures and their off‐target impacts may increase eradication success. With interest in controlling non‐native Poa annua in the region, we aimed to determine if physical and chemical methods can control P. annua (the sub‐Antarctic biotype) in sub‐Antarctic conditions and examined their impact on native plants. We quantified the effectiveness of physical control methods on P. annua in situ on sub‐Antarctic Macquarie Island through field‐based experiments and assessed their selectivity on P. annua compared with native grasses. We also quantified the effectiveness of several herbicides on P. annua at sub‐Antarctic temperatures and assessed their selectivity on native grasses. Of the four physical disturbance methods tested, none effectively reduced P. annua cover as one‐off treatments. Of the herbicide treatments, glyphosate and trifloxysulfuron sodium were effective and were less damaging to native grass species, indicating potential selectivity. Physical control was of limited effectiveness, but did not affect native species richness. An integrated weed management programme utilising the strategic use of selective herbicides with follow‐up chemical and physical intervention may balance control and biodiversity outcomes. This research highlights the importance of site‐specific testing of control methods and understanding off‐target impacts of control when managing alien plant species in protected areas.     

Total mixed ration silage containing elephant grass for small‐scale dairy farms

Alternatives need to be addressed for reducing losses in elephant grass (EG; Pennisetum purpureum) silages. Furthermore, smallholders lack information on the nutritional aspects of dairy farms. The aim of this study was to evaluate total mixed ration silage (TMR) combining fresh EG and concentrate ingredients, creating the following treatments: (i) EG silage (control); (ii) EG, corn and soya bean meal; (iii) EG, corn, soya bean meal and molasses; (iv) EG, citrus pulp and soya bean meal; and (v) EG, citrus pulp, soya bean meal and molasses. Five replicates of each treatment were ensiled in 15‐L plastic jars. The fermentation profile, chemical composition, microbial counts and aerobic stability were assessed. Three contrasts were tested, as follows: (i) control vs. TMR; (ii) TMR with corn vs. TMR with citrus pulp; and (iii) TMR with molasses vs. TMR without molasses. Variables were analysed through the PROC MIXED procedure of the SAS at the 5% level. The TMR had better fermentation profile, lower effluent production and longer aerobic stability compared to control. When the effect of corn was compared to citrus pulp among the TMR, silages with citrus pulp showed lower fermentation losses, non‐protein nitrogen and effluent production. The aerobic stability also improved with citrus pulp. Molasses did not affect the fermentation profile. Overall, lactic acid was the primary acid in all TMR. Nitrogen source (e.g., soya bean meal) can be used without compromising the fermentation process. TMR with citrus pulp showed better results than corn. TMR may be an alternative to optimize the use of EG on smallholdings.     

Determination of Festuca filiformis seedbank characteristics,seedling emergence and herbicide susceptibility to aid management in lowbush blueberry (Vaccinium angustifolium)

Festuca filiformis is a common perennial grass in lowbush blueberry fields, but little is known about the general biology, seedbank characteristics, seedling recruitment or susceptibility of seedlings to currently registered herbicides. The objectives of this research were to determine (i) the presence of F. filiformis seedbanks in lowbush blueberry fields, (ii) whether F. filiformis seedbanks accumulate near the soil surface in lowbush blueberry fields, (iii) the dormancy status of fresh F. filiformis seeds, (iv) the temporal patterns of seedling recruitment in established F. filiformis populations, (v) whether F. filiformis has a vernalisation requirement for flowering in lowbush blueberry and (vi) susceptibility of F. filiformis seedlings to various herbicides currently registered in lowbush blueberry. Festuca filiformis formed a seedbank in lowbush blueberry fields, with an average of 1660 ± 272–5680 ± 1409 seedlings m^?2 emerging from soil cores collected from two infested fields. Most seeds were located at the soil surface, providing opportunities for seedbank management through predation or burning. Fresh seeds lacked dormancy and readily germinated, although germination was reduced by dark conditions. New seedlings emerged in spring and autumn and required vernalisation to flower. Seedlings were susceptible to several currently registered herbicides in lowbush blueberry, although mortality rates were highest in plants treated with glufosinate, flumioxazin, glufosinate + flumioxazin and terbacil. Growers should avoid movement of seeds on machinery, and additional research should be conducted to determine the effects of registered herbicides on F. filiformis seedling recruitment under field conditions in lowbush blueberry.     

草鱼幼鱼肝胰脏抗氧化系统对MC-LR胁迫的响应

为了研究微囊藻毒素(MC-LR)对草鱼(Ctenopharygodon idella)幼鱼肝胰脏抗氧化防御系统的影响,本研究通过腹腔注射的方法(剂量为25、100μg MC-LR·kg-1,分别称为低剂量和高剂量),对草鱼幼鱼进行染毒,于胁迫24、48 h和72 h后分离其肝胰脏,随后采用分光光度法检测了草鱼幼鱼肝胰脏中超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性;同时采用荧光定量PCR方法分析了SOD、CAT、谷胱甘肽过氧化物酶(GPx)和谷胱甘肽还原酶(GR)基因相对表达量的变化。结果显示:高剂量MC-LR诱导24 h和48 h后,草鱼幼鱼肝胰脏SOD活性显著增加(P0.05),而SOD在低剂量组中活性没有显著变化(P0.05);低剂量MC-LR对CAT活性没有显著影响(P0.05),而高剂量MC-LR诱导24 h后可使草鱼幼鱼肝胰脏CAT活性显著增加(P0.05),随后其活性又下降,但差异不显著(P0.05)。在MC-LR胁迫过程中,SOD、CAT、GPx基因表达在两个剂量组中均显著低于对照组(P0.05),而GR基因在低剂量MC-LR胁迫24 h后相对表达量显著上调(P0.05),尽管在72 h相对表达量上调,但差异不显著(P0.05),而高剂量组中GR基因在胁迫24 h和48 h后,其表达量被抑制,但不存在显著差异(P0.05)。进行抗氧化酶活性和基因表达相关性分析发现,SOD和CAT酶活性与SOD和CAT基因表达不相关。上述研究表明MC-LR对草鱼幼鱼肝胰脏抗氧化系统产生了明显的胁迫效应,但MC-LR对机体抗氧化酶活性与基因编码调控之间的相互作用机制还有待更深入的研究。     

陕北黄土区退耕还林(草)地土壤质量特征及其对降水的响应

为了解陕北黄土区退耕还林（草）地土壤质量差异特征及其对降水的响应，该文沿370～470 mm的降水梯度，选取了陕西省吴起县境内的王洼子（370～395 mm）、大吉沟（440～445 mm）和白豹（460～470 mm）3个降水梯度区作为研究区，并结合土壤质量指数法，定量评价降水梯度区广泛栽植的刺槐林（Robinia pseudoacacia）、沙棘林（Hippophae rhamnoides）和草地的土壤物理结构、持水性、盐碱度、碳汇指标以及速效养分指标方面的综合得分，明确植被恢复后土壤质量随降水梯度的变化特征。结果表明：1）降水量、植被类型以及二者的交互作用对土壤指标影响显著（P<0.05）。2）在460～470 mm降水量区，刺槐林（0.829）对土壤质量改良效果最好，其次为沙棘林（0.808），二者土壤质量均达到肥沃水平，且差异不显著，草地（0.679）土壤质量指数最低，土壤为较肥沃水平；在440～445 mm降水量区，沙棘林（0.74）对土壤改良效果最好，其次为刺槐林（0.601），二者均达到较肥沃水平，草地（0.479）土壤质量指数最低为中等水平；在370～395 mm降水量区，3种植被类型土壤质量指数由大到小表现为沙棘林（0.744）>刺槐林（0.387）>草地（0.251），沙棘林土壤质量等级为较肥沃、刺槐林和草地为贫瘠水平。3）在370～470 mm降水梯度内，3种植被类型土壤质量综合指数均随着降水量的减少而降低，其中沙棘林土壤质量指数变异性较低（4.1%），刺槐林（30.1%）和草地（37.2%）土壤质量变异性较高。该研究结果可为陕北黄土区370～470 mm降水梯度下的植被恢复与重建等林业生态工程实施提供数据支撑与理论支持。     

苜蓿-禾草混播对根系可溶性糖含量的影响

可溶性糖对植物的抗冻性存在着积极的影响。为研究苜蓿（Medicago sativa）与禾草混播对根系可溶性糖含量的影响，以苜蓿与无芒雀麦（Bromus inermis Leyss）、披碱草（Elymus dahuricus Turcz）、虉草（Phalaris arundinacea L.）按豆禾间行比例1∶1(B1)、2∶2(B2)、1∶2(B3)和2∶1(B4)混播，并在初霜后测定不同混播处理牧草根系的可溶性糖含量。结果表明，禾草种类对苜蓿根系可溶性糖含量的影响不显著；豆禾间行比例2∶2混播处理苜蓿根系可溶性糖含量显著低于1∶2和2∶1混播处理，1∶1、1∶2和2∶1混播处理间无显著差异；苜蓿与虉草混播后，不同混播比例的虉草根系可溶性糖含量都高于单播虉草。3种禾草中，虉草根系可溶性糖含量显著高于其余两种。综上所述，豆禾间行比为1∶1及1∶2混播处理中苜蓿及禾草的根系可溶性糖含量均较高，考虑苜蓿的耐寒力往往弱于3种禾草，又以间行比1∶2混播的处理为最佳选择。研究为当地苜蓿与禾草混播草地的持续利用提供理论参考。