胭脂鱼仔幼鱼发育及苗种培育技术

对胭脂鱼仔幼鱼发育的形态特征及其主要生活习性进行了描述,探讨了胭脂鱼苗种的不同培育方法,其培育成活率为５５．２％－９７．４％,采用水蝗蚓,鳗鱼饵料,鱼虾蚌肉,动物肝脏及豆制品等多种饵料喂养胭脂鱼鱼种,其综合饵料系数为４．３－５．６,日均增长长度为０．８７－１．０１ｍｍ,日增增生体重为０．０９－０．４５ｇ,全长与全长／体高的关系呈负相关。     

亚蝗微粒子虫毒饵防治东亚飞蝗的研究

用亚蝗微粒子虫毒饵在室内和田间防治东亚飞蝗的试验结果表明，饵料含孢量约１×１０^１２个／ｋｇ，投放量２ｋｇ／６６７ｍ^２，于蝗蝻２龄期施用，１４ｄ内可获得８７．８％的杀蝻效果     

Dietary vitamin C requirement of juvenile Chinese sucker (Myxocyprinus asiaticus)

An 8‐week growth experiment was conducted to quantify the appropriate dietary vitamin C requirement of juvenile Chinese sucker (Myxocyprinus asiaticus). Triplicate groups of 30 experimental fish [initial body weight: (7.1 ± 0.3) g] were cultured in 500 L aquaria and fed with semi‐purified diets containing six levels [10.1 (unsupplemented diet), 37.4, 64.9, 125.2, 244.2 and 482.0 mg kg^?1 diet, respectively] of vitamin C (supplied as L‐ascorbyl‐2‐polyphosphate). Results showed that weight gain of Chinese sucker was significantly increased with increasing dietary vitamin C levels, but there was no significant difference of weight gain among fish fed the diets containing more than 125.2 mg kg^?1 vitamin C. As dietary vitamin C increased, the liver vitamin C content of fish showed the increasing trend firstly and then stabled, while the muscle vitamin C content significantly increased without reaching a constant level. Lower liver malondialdehyde content was observed in 125.2, 244.2 and 482.0 mg kg^?1 vitamin C groups, and higher antioxidant capacity and superoxide dismutase activities were observed in supplemented groups when compared to the unsupplemented group. Liver aspartate aminotransferase and alkaline phosphatase activities were also significant affected by dietary vitamin C. Ash content of fish fed the diet with 244.2 or 482.0 mg kg^?1 vitamin C was significantly higher than that of fish fed the other diets. However, dietary vitamin C had no significant effects on whole‐body crude protein, lipid and moisture contents. The vitamin C requirement of juvenile Chinese sucker was estimated to be 84.6 and 126.1 mg kg^?1 based on weight gain and liver vitamin C concentration respectively.     

Hatching rate of Chinese sucker (Myxocyprinus asiaticus Bleeker) eggs exposed to total dissolved gas (TDG) supersaturation and the tolerance of juveniles to the interaction of TDG supersaturation and suspended sediment

Flood discharge containing excessive suspended sediments (SS) can result in total dissolved gas (TDG) supersaturation downstream of dams. It has been confirmed that high TDG levels and excessive SS cause serious threats to fish survival. To explore the effects of TDG supersaturation on the hatchability of Chinese sucker (Myxocyprinus asiaticus Bleeker) eggs, eggs were exposed to 100%, 125%, 130% and 135% TDG supersaturated water. The results showed that the hatching rates were 78.25%, 75.35%, 71.30% and 67.50% at the 100%, 125%, 130% and 135% TDG respectively. The hatching rate decreased with increasing TDG levels. Furthermore, acute lethality experiments were carried out to investigate the interaction of TDG supersaturation and SS on Chinese sucker. Half‐year‐old juveniles were exposed to TDG supersaturated water with SS concentrations of 0, 200, 600 and 1,000 mg/L, and the TDG levels were 125%, 130%, 135% and 140% respectively. The fish exhibited abnormal behaviours and signs of gas bubble disease. Mortality increased with increasing TDG levels and SS concentrations. The Chinese suckers only survived for 2.5 hr at 140% TDG and 1,000 mg/L SS. Moreover, an increase in SS in TDG supersaturated water could cause a decrease in the median lethal time (LT₅₀). When the TDG level reached 140%, the LT₅₀ declines significantly at 600 and 1,000 mg/L SS. The interaction of TDG supersaturation and SS was significant at LT₅₀ (F = 22.81, df = 9, p < 0.001). The results indicate that the combined effect of TDG and SS could be great on Chinese sucker.     

胭脂鱼仔鱼的生长和消化酶活力变化

对胭脂鱼(Myxocyprinus asiaticus)孵化后1～37 d仔鱼的体长、体重、RNA/DNA比率、胰蛋白酶、脂肪酶和淀粉酶的酶活力变化进行了测定。结果显示:胭脂鱼仔鱼1～37 d体长稳定增长。体重增加呈阶段性,在1～21 d体重增长缓慢,21～29 d体重增加较快,29～37 d体重快速增加。仔鱼从7 d开始摄食,1～6 d,RNA/DNA比率变化很小,蛋白含量逐渐降低;7～21 d,RNA/DNA比率波动较大,蛋白含量增长缓慢;21～37 d,RNA/DNA比率和蛋白质均增长迅速。生长率(GR)与RNA/DNA比率的回归方程为:GR=1.01RNA/DNA+1.6077(R2=0.8516)。从2、5和7 d开始分别检测到明显的胰蛋白酶、脂肪酶和淀粉酶活力,之后,消化酶活力随着发育先快速地上升然后波动性地下降到一个稳定水平。结果表明:胭脂鱼仔鱼从内源性到外源性营养转化过程中,不同的消化酶能够检测到明显酶活力的时间不同,到仔鱼开始摄食时已经具备消化蛋白质、脂肪和淀粉的能力;RNA/DNA比率可以作为判定仔鱼生长率的可靠指标。     

饲料脂肪水平对胭脂鱼幼鱼生长、体组成和抗氧化能力的影响

试验选用体重(6.73±0.21)g的胭脂鱼(Myxocyprinus asiaticus)幼鱼540尾,随机分成6个组,每组3个重复,每个重复30尾试验鱼。以大豆油为脂肪源,配制成脂肪水平为2.04%、4.43%、6.88%、9.02%、11.98%、13.39%的半精制饲料,进行为期56 d的生长试验,研究脂肪水平对胭脂鱼幼鱼生长、体组成及机体抗氧化能力的影响。结果显示:饲料脂肪水平对胭脂鱼的生长性能有显著影响(P<0.05)。脂肪水平为6.88%时,胭脂鱼增重率、特定生长率、蛋白质沉积效率最高,饲料系数最低。脂肪沉积效率随脂肪水平的增加呈下降趋势。随脂肪水平增加,胭脂鱼全鱼、肌肉和肝胰脏脂肪含量逐渐上升,至6.88%组后稳定,而全鱼水分、粗蛋白含量没有显著变化(P>0.05)。胭脂鱼机体抗氧化能力随着脂肪水平的增加呈现先升后降趋势,脂肪水平为6.88%时,总抗氧化能力(T-AOC)、超氧物歧化酶(SOD)值最大,丙二醛(MDA)值最小,即抗氧化能力最强。以增重率、特定生长率和饲料系数进行回归分析并考虑脂肪水平对胭脂鱼营养组成和抗氧化能力的影响,确定胭脂鱼幼鱼适宜脂肪水平为6.62%~7.02%。     

Detection of ‘Candidatus Liberibacter asiaticus’ in citrus by concurrent tissue print-based qPCR and immunoassay

‘Candidatus Liberibacter asiaticus’ (CLas) is associated with the most destructive disease of citrus, huanglongbing (HLB). The most widely used methods for detection of CLas are PCR-based and require purification of DNA from plant samples. Elution of DNA from tissue prints made on nitrocellulose membranes followed by qPCR (TPE-qPCR) was compared to DNA extraction of plant tissue followed by PCR (X-PCR) by testing the same tissue samples. The former estimated a higher CLas population in tissue prints than the latter (t-test; P = 0.009). All extracts prepared for TPE-qPCR throughout the experiment were also tested by conventional PCR and 80.8% were identified as positive. A similar set of stem and petiole tissue samples was tested by TPE-qPCR and immunoassay. Although the detection rate by TPE-qPCR was higher than by immunoassay, about 6% of tissue prints were positive by immunoassay but not by TPE-qPCR. Thus, a higher detection rate would be achieved by combining TPE-qPCR with immunoassay. Significant differences were observed in the performance of nitrocellulose membranes from different manufacturers in these assays. Immunotissue prints showed that the spatial distribution pattern of CLas infection varied widely from one sample to another, but the patterns were highly correlated among serial sections from the same sample, suggesting that CLas preferentially colonizes adjacent phloem cells in a vertical rather than horizontal direction.     

柑橘黄龙病菌在染病贡柑枝条和果实橘络内的分布规律

 柑橘黄龙病是柑橘生产上最具毁灭性的病害之一,由目前还无法分离培养的候选韧皮部杆菌亚洲种(“Candidatus Liberibacter asiaticus”, CLas)引起的。早期研究表明CLas在染病柑橘植株内的分布并不均匀。为进一步探究CLas在染病枝条及果实内的空间分布规律,本研究以感染黄龙病的贡柑(带果实)枝条为材料,利用定量PCR(quantitative PCR, qPCR)分析同一枝条不同部位及果实橘络中的CLas浓度。结果表明,带果贡柑枝条不同部位的CLas分布不均匀且以果实橘络部位的CLas浓度最高(15 487.6 CLas cells/ng总DNA)。通过对同一橘络不同片段长度(每1.5 cm)中的CLas进行定量分析发现,同一橘络中的CLas同样呈不均匀分布(CLas个数范围:12 407~10 271 089)。此外,定量分析发现黄龙病引起的畸形果大小两侧橘络中的CLas浓度差异不显著。该结果可为探究CLas在柑橘枝条及果实中的转运规律提供参考。     

马蜂柑响应黄龙病菌不同侵染时期的生物学和转录组学分析

【目的】 柑橘黄龙病（Huanglongbing,HLB）是制约柑橘产业发展的重大病害,我国发生的黄龙病由韧皮部杆菌亚洲种（‘Candidatus Liberibacter asiaticus’,CLas）引起。本研究通过分析潜在耐黄龙病柑橘品种马蜂柑（Citrus hystrix）在感染黄龙病后不同时期的生物学症状、显微结构和转录组学差异,揭示马蜂柑不同时期对黄龙病菌具有耐性的分子机理,为进一步深入挖掘抗性基因及开展抗病育种打下基础。 【方法】 以嫁接在两年生卡里佐枳橙砧木上的马蜂柑作为供试材料,使用只感染CLas、其他常见柑橘病毒类病原均呈阴性的毒源为接毒材料对马蜂柑进行接种,并在接种毒源后每隔15 d进行定期的荧光定量PCR检测。将马蜂柑接种毒源4个月（最早检测到CLas阳性）作为感病前期处理,接种毒源14个月作为感病后期处理,以健康植株为对照,进行生物学症状和显微结构观察,分析其感病后不同时期的结构变化。结合比较转录组学分析与荧光定量PCR验证,解析其耐病相关机制。 【结果】 症状观察发现,马蜂柑在感病前期和感病后期,植株叶片几乎不显症;显微结构观察表明,马蜂柑在感病前期中脉组织结构清晰,细胞形态正常,无淀粉粒积累的现象,仅在后期出现韧皮部极少数的筛管被堵塞;对转录组测序结果进行筛选鉴定,马蜂柑感病前期共筛选鉴定到181个差异表达基因,感病后期共筛选鉴定到1 384个差异表达基因;比较转录组分析表明,马蜂柑感病前期和后期的差异表达基因主要涉及细胞壁代谢、防御反应、淀粉与蔗糖代谢、胼胝质合成以及信号转导等方面,其中在感病前期,淀粉与蔗糖代谢、细胞壁代谢相关的调控基因下调表达,在感病后期,水杨酸代谢及其信号转导途径、病程相关蛋白和谷胱甘肽转移酶相关的调控基因上调表达。 【结论】 马蜂柑响应CLas早期侵染主要表现为物理结构稳定、淀粉合成和光合作用等途径不受干扰;而水杨酸介导的抗性信号转导、效应蛋白触发免疫反应（effector-triggered immunity,ETI）激活的病程相关蛋白和谷胱甘肽转移酶参与的解毒作用是感病后期的主要耐病机制。     

胭脂鱼苗种工厂化繁育技术研究

胭脂鱼苗种工厂化繁育技术研究结果表明,运用智能控温及水处理技术,将水温准确控制到(18±1)℃,亲本通过强化培育、人工催熟和催情;使产卵率达100%、受精率达95%,孵化率达93%。2009—2011年共繁殖鱼苗2 607万尾、培育鱼种1 879万尾。