生物工程应用于作物育种的试验研究 第4报 外源DNA处理水稻遗传变异的初步研究

以外源牛胰 DNA 培养处理水稻萌动种子,研究 D_1、D_2和 D_3植株的形态特征和生理特性的遗传变异。结果表明,外源 DNA 处理所引起的变异类型多、幅度大、频率高,但与外源 DNA 浓度之间没有明显相关.外源 DNA 处理对改良现有推广品种的某一不良性状较为有利,同时在变异后代中可获得优良的或特殊的变异类型.     

2BD-7型多功能精密耕播机的研制与试验研究

2BD-7型多功能精密耕播机是与大豆“暗垄密”高产栽培技术相配套的播种机械,该机施肥采用滑刀式开沟器,可实现种、底肥的分层施撒;播种采用双条精量点播,可一次完成开沟、施肥、播种、覆土工序。生产试验表明,各行排量一致性、总排量稳定性均达到国标对播种大豆的播种质量指标。各项技术性能满足大豆“暗垄密”高产栽培技术所提出的农艺要求。     

传统池塘和“跑道”养殖黄金鲫的营养组成分析与评价

为研究不同养殖模式对黄金鲫(Carassius auratus)营养组成和品质的影响,以传统池塘精养和采用池塘内循环流水“跑道”模式养成的黄金鲫为研究对象,分析比较了两种养殖模式下黄金鲫肌肉营养成分、质构和风味物质等指标。结果显示,“跑道”组黄金鲫背肌的粗脂肪含量显著低于池塘组,而粗灰分含量显著高于池塘组(P＜0.05);“跑道”组黄金鲫肌肉的硬度、粘附性和黏性等质构指标与池塘组相比显著提升(P＜0.05)。两种模式下养成的黄金鲫,其肌肉的氨基酸组成无显著差异(P＞0.05),且必需氨基酸评分均高于FAO/WHO的标准。“跑道”组黄金鲫肌肉中不饱和脂肪酸、单不饱和脂肪酸和多不饱和脂肪酸占总脂肪酸的比例均显著高于池塘组,而饱和脂肪酸占比显著低于池塘组(P＜0.05)。“跑道”组黄金鲫肌肉中部分挥发性物质,如壬醛、1-辛烯-3-醇的含量也低于池塘组。分析比较结果表明,采用池塘内循环流水“跑道”养殖模式可以在一定程度上提升黄金鲫的营养品质。     

太平洋牡蛎(Crassostrea gigas)类α-1,2-岩藻糖基转移酶的密码子优化与原核表达

牡蛎消化组织内存在的类A型血型组织抗原是其特异性富集诺如病毒的主要原因,FUT2(Fucosyltransferase 2,α-1,2-岩藻糖基转移酶)是A型血型组织抗原合成的关键酶.本研究在前期克隆了太平洋牡蛎(Crassostrea gigas)类FUT2基因cDNA全长的基础上,根据大肠杆菌密码子偏爱性优化并合成了类FUT2基因,插入原核表达载体pRSET A构建pRSET-mof,将其转化大肠杆菌BL21,用异丙基-β-D-硫代半乳糖苷(IPTG)诱导.经SDS-PAGE分析显示,在37℃、IPTG终浓度为0.8 mmol/L的条件下,诱导4h后出现大小约为46 kDa的特异性目的条带.利用His亲和层析柱纯化及超滤管浓缩目的蛋白,得到单一条带,说明纯化效果良好.Western blot分析显示,目的蛋白与抗6×His标签单克隆抗体、抗人FUT2单克隆抗体均能发生特异性反应,表明优化后的太平洋牡蛎类FUT2基因在大肠杆菌系统中成功表达.本研究结果为今后研究太平洋牡蛎类FUT2基因的功能,进一步探索牡蛎特异性富集诺如病毒的分子机理奠定了基础.     

Effect of dietary <Emphasis Type="SmallCaps">l</Emphasis>-tryptophan on cannibalism,survival and growth in pikeperch <Emphasis Type="Italic">Sander lucioperca</Emphasis> (L.) post-larvae

The effect of supplemented commercial diets with crystalline l-tryptophan (TRP—5, 10 and 20 g TRP kg^?1) on cannibalism, survival and growth parameters of pikeperch post-larvae (Sander lucioperca) was evaluated. Fifteen-day-old pikeperch larvae (mean weight—6.8 mg) were reared during the next 28 days (20.5 °C, 16L:8D) in glass aquaria in a recirculating aquaculture system. The enzyme-linked immunosorbent assay showed that TRP-supplemented diets were effective in increasing the levels of serotonin (5-HT) in the body tissue of pikeperch. TRP supplementation resulted in a slight decrease in both types of cannibalism, although the reduction in cannibalism impact did not amount to more than a few percent. TRP treatment had no significant influence on the final survival of pikeperch post-larvae (ranged from 20.1 ± 12.4 to 29.0 ± 12.9 %). However, contrary to the earlier studies conducted on other fish species, no significant difference in the growth rates and feeding behavior of pikeperch between TRP-fed and control group were found. The final body weight and growth rate ranged from 0.211 ± 0.014 to 0.243 ± 0.016 and from 12.19 ± 0.38 to 12.76 ± 0.35 % day^?1, respectively. To our knowledge, this is the first study reporting the effects of TRP supplementation on the cannibalism–survival–growth relations in fish in the post-larval stage.     

Effects of nucleotides on growth performance,immune response,disease resistance and intestinal morphology in shrimp <Emphasis Type="Italic">Litopenaeus vannamei</Emphasis> fed with a low fish meal diet

Nucleotides (NT) are widely used as an immunostimulant in aquaculture. A 10-week feeding trial was conducted to evaluate the effects of dietary NT on the growth performance, immune response, disease resistance and intestinal morphology of the Pacific white shrimp Litopenaeus vannamei (initial mean body weight: 0.39 ± 0.00 g). Five isolipidic (about 7 % crude lipid) and isonitrogenous (about 39 % crude protein) practical diets were supplemented with graded levels of NT (0, 60, 90, 120 and 1200 mg kg^?1), respectively. These diets were named as N1, N2, N3, N4 and N5. Each diet was randomly fed to six tanks of shrimps, and each tank contained 40 shrimps. The shrimps were fed four times daily (07:00, 11:00, 16:00 and 21:00 h). The results showed that there were no significant differences in survival, final weight, specific growth rate, feed intake, feed conversion ratio and whole-body compositions among all the treatments (P > 0.05). Shrimps fed the control diet had the significantly lowest activities of superoxide dismutase, total nitric oxide synthase and lysozyme (P < 0.05). However, those parameters increased when dietary NT increased from 60 to 120 mg kg^?1 and decreased as inclusion level increased to 1200 mg kg^?1 (P < 0.05). The cumulative mortality of the shrimps challenged with Vibrio parahaemolyticus was significantly higher in the treatments without dietary NT supplementation than those in treatments with dietary NT (≥90 mg kg^?1) (P < 0.05). The jejunum wall thickness in shrimps fed the control diet was significantly lower than that in the treatments of N2, N3 and N4 (P < 0.05). For the villus height, the highest value (49.29 µm) was found in N3. In summary, the present study showed that 90 mg kg^?1 of dietary NT is the optimum dietary level for good gut health, immune response and disease resistance of Pacific white shrimp fed a diet with 18 % fish meal.     

Performance of <Emphasis Type="Italic">Litopenaeus vannamei</Emphasis> postlarvae reared in indoor nursery tanks under biofloc conditions at different salinities and zero-water exchange

Shrimp farming at low salinity is a trend that will continue to grow globally. Performance of Litopenaeus vannamei postlarvae in the nursery at different salinities with a biofloc technology (BFT) system needs to be explored further, as the nursery is currently used as a transitional stage between the hatchery and grow-out ponds. Hence, this study evaluated the effect of seven salinity levels (2, 4, 8, 12, 16, 25, and 35 ‰) on the performance of L. vannamei postlarvae reared with a BFT system and zero-water exchange at 2000 org/m³. Additionally, this study evaluated the water quality of all salinity treatments. After 28 days of culture, the findings showed that, under biofloc conditions, salinity affected the performance of some variables of water quality in some cases, but only the combination of a high nitrite-N concentration (>4 mg/l) and low salinity (2 and 4 ‰) caused up to 100 % shrimp mortality in the first 2 weeks. In the rest of the treatments (8, 12, 16, 25, and 35  ‰), shrimp survival was >72 %. Shrimp mortality was affected by salinity, especially when it decreased from 35–25–16 to 12 and 8 ‰. The organisms reared at low salinities presented lower final weights and specific growth rate than those reared at higher salinities. An inverse relationship was shown between the ion concentration and the final weight of shrimp.     

Effects of potential probiotic <Emphasis Type="Italic">Lactococcus lactis</Emphasis> subsp. <Emphasis Type="Italic">lactis</Emphasis> on digestive enzymatic activities of live feed <Emphasis Type="Italic">Artemia franciscana</Emphasis>

In the present study, cellulase, protease, lipase and amylase activities were performed to investigate the effects of Lactococcus lactis subsp. lactis CF4MRS bioencapsulation of Artemia franciscana. Our results show that cellulase activities (total cellulase—FPase activity, exoglucanase and endoglucanase—CMCase activity) were significantly higher (P < 0.05) in A. franciscana tissue homogenates compared to those in the control group after 8 h of L. lactic bioencapsulation. Notably, an exception case was found in β-D-glucosidase activity, whereby the cellulase activity was not significantly different (P > 0.05) compared to the control group. Administrations of L. lactis at cell concentration of 10⁸ CFU mL^?1 showed considerable improvement of other important enzymatic activities such as amylase, protease and lipase in A. franciscana. The amylase/protease ratio in probiotic-treated A. franciscana was recorded at 0.343, approximately two times higher than those without probiotic administration (0.184). In contrary, amylase/lipase ratio showed half of a reduction (0.330) in L. lactis-administrated A. franciscana compared to the control (0.614). Our study suggests that important digestive enzymes, e.g., cellulase, amylase, protease and lipase, can be enhanced through bioencapsulation of A. franciscana with L. lactis subsp. lactis, which could in turn lead to further stimulation of endogenous enzymes in the fish and shrimp larvae.     

Effect of the early temperature on the growth of larvae and postlarvae turbot, <Emphasis Type="Italic">Scophthalmus maximus</Emphasis> L.: muscle structural and ultrastructural study

Turbot specimens were kept at three temperatures (T _s ): warm (W) (21–22 °C), ambient (A) (17–18 °C) and cold (C) (13–14 °C) during the larval and early postlarval stages. At 90 days posthatching (dph), all of them were transferred to ambient T until 190 dph. At 2–3 dph, the specimens showed a monolayer of red muscle and immature white fibres; external or dermomyotome cells (presumptive myogenic cells) were observed on the surface of the red muscle. In the following stages, many myogenic cells and presumptive myogenic precursors were observed within the myotome, presumably derived of the dermomyotome. When comparing the growth at the same age (2, 10, 25, 37 dph), the body length and the muscle growth were positively influenced by the warm T, being the hyperplasia the muscle parameter more significantly influenced. The development rate was also positively correlated with the high T: the beginning of the metamorphosis took place at 15, 23 and 25 dph at W, A and C temperatures, respectively, with the highest body length values at ambient temperature. The metamorphosis finished at 25, 30 and 37 dph at W, A and C temperatures, respectively, with the highest body length values at warm temperature. However, the muscle cellularity was similar in all the groups at the end of the metamorphosis. At 90 and 190 dph, the largest body length was observed at W temperature. However, the muscle cellularity was similar between A and W; the number of fibres was similar in all the groups at 190 dph, which shows the beginning of a compensatory muscle growth in A and C, mainly in A.     

Astaxanthin degradation and lipid oxidation of Pacific white shrimp oil: kinetics study and stability as affected by storage conditions

The kinetics of astaxanthin degradation and lipid oxidation in shrimp oil from hepatopancreas of Pacific white shrimp (Litopenaeus vannamei) as affected by storage temperature were studied. When shrimp oil was incubated at different temperatures (4, 30, 45 and 60 °C) for 16 h, the rate constants (k) of astaxanthin degradation and lipid oxidation in shrimp oil increased with increasing temperatures (p < 0.05). Thus, astaxanthin degradation and lipid oxidation in shrimp oil were augmented at high temperature. When shrimp oils with different storage conditions (illumination, oxygen availability and temperature) were stored for up to 40 days, astaxanthin contents in all samples decreased throughout storage (p < 0.05). All factors were able to enhance astaxanthin degradation during 40 days of storage. With increasing storage time, the progressive formation of primary and secondary oxidation products were found in all samples as evidenced by the increases in both peroxide values (PV) and thiobarbituric acid reactive substances (TBARS) (p < 0.05). Light, air and temperatures therefore had the marked effect on astaxanthin degradation and lipid oxidation in shrimp oils during the extended storage.