草鱼呼肠孤病毒VP6蛋白在毕赤酵母中表达的初步研究

根据GeneBank中草鱼呼肠孤病毒(grass carp reovirus,GCRV)104株VP6蛋白的全基因序列(HM234682)设计一对特异引物,以提取的GCRV-104核酸为模板,采用RT-PCR技术扩增VP6蛋白编码基因,并将其克隆到含强启动子AOXⅠ的毕赤酵母(Pichia pastoris)表达载体pPICZB上。重组酵母质粒pPICZB-VP6经SacⅠ线性化,电击转化到毕赤酵母KM71菌株中,Zeocin抗性平板上筛选高拷贝转化子。阳性转化子在30℃,0.5%(V/V)的甲醇添加量的条件下,连续诱导3 d,表达产物经SDS-PAGE和Western-Blotting检测,结果表明成功实现了GCRV-104 VP6蛋白在毕赤酵母中的胞内表达,重组VP6蛋白相对分子质量约46 000,与天然VP6蛋白相对分子质量接近,并且可与鼠抗草鱼GCRV-104 VP6蛋白的多克隆抗体特异性结合。     

二倍体和四倍体泥鳅线粒体ND-5/6基因多态性的PCR-RFLP分析

采用PCR-RFLP分析方法,对洞庭湖、武汉两地的二倍体和四倍体泥鳅线粒体DNAND-5/6基因多态性及4个群体遗传变异和遗传关系进行了研究。结果表明,120尾泥鳅mtDNAND-5/6扩增片段长度均为2.2kb;从13种限制性内切核酸酶中筛选出6种多态性内切酶(HaeⅢ、DraⅠ、RsaⅠ、TaqⅠ、HinfⅠ、MspⅠ),对扩增产物进行酶切,共检测到66种单倍型。泥鳅群体内单倍型多样性和核苷酸多样性分别为0.7679～0.9385和0.01041～0.03212;其中,洞庭湖二倍体核苷酸多样性最高(0.03212),武汉二倍体其次(0.02452),二倍体群体内核苷酸多样性高于四倍体。群体间的核苷酸多样性(π)大小为0.02588～0.04144,平均值为(0.031737±0.000005)。群体间的核苷酸歧化距离(δ)大小为0.00462～0.01617,平均值为(0.010659±0.000003);其中,武汉二倍体和四倍体之间的核苷酸歧化距离最大(0.01617),武汉四倍体和洞庭湖二倍体之间的核苷酸歧化距离最小(0.00462)。MonteCarlo模拟x2检验表明,4个群体间的单倍型频率存在极...     

Optimal chemical triploid induction in geoduck clams, Panopea abrupta, by 6-dimethylaminopurine

The development of geoduck culture techniques coupled with increased market demand during the past decade led to cultured intertidal geoduck beds, over which concerns have arisen regarding the potential genetic risk posed by the reproductive contribution of hatchery outplants to wild stocks. Although an ongoing study to determine the genetic stock structure of Puget Sound geoducks has yet to yield definitive results, sterile triploids would enable geoduck culture to proceed irrespective of any genetic differences found, and may increase rate of growth. At 15 °C and salinity of 30‰, we evaluated the suitability of cytochalasin B (CB) and 6-dimethylaminopurine (6-DMAP) for triploid induction in geoducks. Parallel tests of induction agent, start time, duration, and concentration were conducted on the same gamete pool. We found optimal triploid induction (92%) and suitable survivorship (30%) resulted from a 600 μM 6-DMAP treatment. This work enables the future examination of the efficacy of triploid induction for conferring sterility and increasing growth rate in this commercially valuable species.     

Effect of dietary vitamin B6 supplementation on growth and intestinal microflora of juvenile golden pompano (Trachinotus ovatus)

In order to investigate the dietary vitamin B₆ (VB₆) requirement for juvenile golden pompano, an experiment of six different diets with six dietary VB₆ levels (0, 3.75, 7.47, 10.6, 13.7 and 18.5 mg/kg) was conducted. The results indicated that the content of dietary VB₆ significantly increased weight gain rate (WGR), specific growth rate (SGR) and feed conversion ratio (FCR; p < 0.05). Golden pompano fed diet containing 7.47 mg/kg VB₆ achieved the maximum ALT, AST, GPx, hepatic VB₆ concentrations as well as the minimum MDA, however, the ALT, AST, GPx and hepatic VB₆ concentrations decreased while MDA increased with a further increase in dieatry VB₆. The highest values of GR and POD occurred at the 10.6 and 13.7 mg/kg dietary VB₆ (p < 0.05) respectively. A diet supplemented with 7.47 mg/kg VB₆ increased intestinal Na⁺, K⁺‐ATPase, Chymotrypsin, γ‐glutamyl transpeptidase, creatine kinase and amylase activities (p < 0.05). The relative abundance of Tenericutes and Bacteroidetes decreased while Proteobacteria and Firmicutes improved with an increase in dietary VB₆ levels up to 7.47 mg/kg. Quadratic regression analysis on WGR, hepatic VB₆ concentrations, AST and ALT indicated that the optimum dietary VB₆ levels for juvenile golden pompano were 8.84–9.28 mg/kg.     

Effect of crude oil extracts from trout offal as a replacement for fish oil in the diets of the Australian native fish Murray cod Maccullochella peelii peelii

The efficacy of trout oil (TO), extracted from trout offal from the aquaculture industry, was evaluated in juvenile Murray cod Maccullochella peelii peelii (25.4±0.81 g) diets in an experiment conducted over 60 days at 23.7±0.8 °C. Five isonitrogenous (48% protein), isolipidic (16%) and isoenergetic (21.8 kJ g^?1) diets, in which the fish oil fraction was replaced in increments of 25% (0–100%), were used. The best growth and feed efficiency was observed in fish fed diets containing 50–75% TO. The relationship of specific growth rate (SGR), food conversion ratio (FCR) and protein efficiency ratio (PER) to the amount of TO in the diets was described in each case by second‐order polynomial equations (P<0.05), which were: SGR=–0.44TO²+0.52TO+1.23 (r²=0.90, P<0.05); FCR=0.53TO²–0.64TO+1.21 (r²=0.95, P<0.05); and PER=–0.73TO²+0.90TO+1.54 (r²=0.90, P<0.05). Significant differences in carcass and muscle proximate compositions were noted among the different dietary treatments. Less lipid was found in muscle than in carcass. The fatty acids found in highest amounts in Murray cod, irrespective of the dietary treatment, were palmitic acid (16:0), oleic acid (18:1n‐9), linoleic acid (18:2n‐6) and eicosapentaenoic acid (20:5n‐3). The fatty acid composition of the muscle reflected that of the diets. Both the n‐6 fatty acid content and the n‐3 to n‐6 ratio were significantly (P<0.05) related to growth parameters, the relationships being as follows. Percentage of n‐6 in diet (X) to SGR and FCR: SGR=–0.12X²+3.96X–32.51 (r²=0.96) and FCR=0.13X²–4.47X+39.39 (r²=0.98); and n‐3:n‐6 ratio (Z) to SGR, FCR, PER: SGR=–2.02Z²+5.01Z–1.74 (r²=0.88), FCR=2.31Z²–5.70Z+4.54 (r²=0.93) and PER=–3.12Z²–7.56Z+2.80 (r²=0.88) respectively. It is evident from this study that TO could be used effectively in Murray cod diets, and that an n‐3:n‐6 ratio of 1.2 results in the best growth performance in Murray cod.     

Regeneration of adventitious shoots from mature stored cotyledons of Japanese plum (Prunus salicina Lind1)

Adventitious shoot regeneration from mature stored cotyledons of Japanese plum (Prunus salicina Lind1) was achieved in vitro. The influences of the presence and absence of the light, different concentrations of thidiazuron (TDZ) and benzyladenine (BA) in the culture media, TDZ pretreatments and different basal salts on shoot regeneration were evaluated. TDZ was more effective in inducing shoot regeneration from mature stored cotyledons than BA. Dark incubation significantly increased the regeneration frequencies. Quoirin/Lepoivre (QL) basal salts stimulated shoot regeneration more than woody plant (WPM) or B5 salts did. The frequency of adventitious shoot formation varied among the varieties and the regeneration ability appeared to be genotype depended. The frequency of regeneration under the optimum tested conditions for ‘Bruce’, ‘Shiro’, ‘Redheart’, ‘Gladstone’ and ‘Early Golden’ cotyledons were 66.7%, 46.7%, 43.3%, 26.7% and 6.7%, respectively.     

In vitro high frequency direct plant regeneration from whole leaves of blackberry

High frequency and direct (without callus) plant regeneration was achieved from whole leaf explants of thornless blackberry (Rubus hybrid) cv. Black Satin (EC No. 381258; PI No. 553272) in vitro. Leaf blade explants from 1-, 3- and 5-month-old mother cultures were cultured on Murashige and Skoog (MS) medium with thidiazuron (TDZ), N⁶-benzylaminopurine (BAP), indol-3-butyric acid (IBA) and α-naphthalene acetic acid (NAA), alone or in combination. Three-month explants cultured on 0.02 mg l⁻¹ TDZ produced a high regeneration frequency (91.7%) and the most shoots/leaf explant (17.3). The shoot primordia developed within 3 weeks from the point of detachment of the petiole from the leaf blade. The age of the explant source significantly affected the shoot regeneration potential of the leaf explants. Leaves excised from 3-month-old in vitro-cultured shoots performed better than those from 1- and 5-month-old shoots. Shoots rooted best on half-strength MS basal medium with 0.5 mg l⁻¹ IBA and 90% of the plantlets survived acclimatization. The regenerated plantlets were morphologically similar to the mother plants.     

Floral induction (FI) in longan (Dimocarpus longan, Lour.) trees. III: Effect of shading the trees on potassium chlorate induced FI and resulting hormonal changes in leaves and shoots

Previous experiments demonstrated that treatment of longan trees with potassium chlorate (KClO₃) induces “off season floral induction” (FI) even in the absence of the naturally required cool temperature [Manochai, P., Sruamsiri, P., Wiriya-alongkorn, W., Naphrom, D., Hegele, M., Bangerth, F., 2005. Year around off season flower induction in longan (Dimocarpus longan, Lour.) trees by KClO₃ applications: potentials and problems. Sci. Hortic. 104, 379–390]. Potassium chlorate, however, cannot replace the presence of functional mature leaves and sufficient light intensity. Here we examined in more detail the effect of shade (about 10% of natural sunlight) on KClO₃ affected hormone concentrations/transport of leaves and shoot apical buds (SAB) and their interactions with FI.     

茄子新品种新茄6号的选育

新茄6号是由从新疆长茄中单株套袋自交选育出的自交系4号长茄和从前苏联长茄品种中单株套袋自交选育出的自交系26号长 茄配组,然后在后代分离群体中通过系统选择育成的长茄新品种。植株生长势强,第1花生长在第5~6片叶腋,果实细长,浅紫红色, 光泽度好,平均单果质量140 g,抗黄萎病能力优于新疆长茄,一般每667 m²产量4 500~5 500 kg,适于新疆早春大棚和露地覆膜早 熟栽培。