烟草线条病毒RT-PCR检测方法

根据已报道的烟草线条病毒外壳蛋白基因序列，设计出一对特异性引物，提取大豆病叶的总RNA，反转录成cDNA，PCR扩增出约400bp大小的产物，产物克隆到pGEM-T Easy载体上，转化大肠杆菌DH 5α，抽提的质粒用Not Ⅰ酶切，出现430bp大小的目标插入片段。经序列测定确认了插入片段的大小为404bp。     

C/N ratio control and substrate addition for periphyton development jointly enhance freshwater prawn Macrobrachium rosenbergii production in ponds

The present research investigated the effect of carbon/nitrogen ratio (C/N ratio) control in ponds with or without substrate addition for periphyton development on production of giant freshwater prawn. C/N ratios of 10, 15 and 20 were investigated in 40 m^− 2 ponds stocked with 2 prawn juveniles (5.023 ± 0.02 g) m^− 2 with or without added substrates for periphyton development. The various treatment combinations of C/N ratio and periphyton substrate addition are abbreviated as ‘CN10’, ‘CN15’, ‘CN20’, ‘CN10 + P’, ‘CN15 + P’ and ‘CN20 + P’, P representing periphyton substrate. A locally formulated and prepared feed containing 30% crude protein with C/N ratio10 was applied. Tapioca starch was used as carbohydrate source for manipulating C/N ratio and applied to the water column separately from the feed. Increasing the C/N ratio from 10 to 20 reduced (P < 0.001) the total ammonia-nitrogen (TAN), nitrite–nitrogen (NO₂–N) and nitrate–nitrogen (NO₃–N) in water column and total Kjeldahl nitrogen (TKN) in sediment. The addition of substrates only influenced the NO₂–N concentration in the water column (P < 0.001). Increasing the C/N ratio raised the total heterotrophic bacterial (THB) population in the water column, sediment and periphyton (P < 0.001). It also increased the dry matter (DM), ash free dry matter (AFDM), and chlorophyll a content of periphyton (P < 0.001). The lowest specific growth rate (SGR), the highest food conversion ratio (FCR), and the lowest protein efficiency ratio (PER) were recorded in treatment CN10 (P < 0.05). The addition of substrates did not influence size at harvest (P > 0.05) but improved the survival from 62.8 to 72% (P < 0.001). Increasing the C/N ratio from 10 to 20 increased the net yield by 40% and addition of substrate increased the net yield by 23%. The combination of C/N ratio control and substrate addition increased the net yield by 75% from 309 (CN10) to 540 (CN20 + P) kg ha^− 1 (120 days)^− 1. This 75% higher production concurred with (1) a lower inorganic nitrogen content in the water column, (2) a higher THB abundance supplying additional single cell protein to augment the prawn production, and (3) an improved periphyton productivity and quality.     

Pigeonpea improvement: An amalgam of breeding and genomic research

In the past five decades, constant research has been directed towards yield improvement in pigeonpea resulting in the deployment of several commercially acceptable cultivars in India. Though, the genesis of hybrid technology, the biggest breakthrough, enigma of stagnant productivity still remains unsolved. To sort this productivity disparity, genomic research along with conventional breeding was successfully initiated at ICRISAT. It endowed ample genomic resource providing insight in the pigeonpea genome combating production constraints in a precise and speedy manner. The availability of the draft genome sequence with a large‐scale marker resource, oriented the research towards trait mapping for flowering time, determinacy, fertility restoration, yield attributing traits and photo‐insensitivity. Defined core and mini‐core collection, still eased the pigeonpea breeding being accessible for existing genetic diversity and developing stress resistance. Modern genomic tools like next‐generation sequencing, genome‐wide selection helping in the appraisal of selection efficiency is leading towards next‐generation breeding, an awaited milestone in pigeonpea genetic enhancement. This paper emphasizes the ongoing genetic improvement in pigeonpea with an amalgam of conventional breeding as well as genomic research.     

A 3-Year Adaptation Study of Three Distinct Grapevine Cultivars under Midwestern Field Conditions

Simultaneous responses of American (Concord), European (Pinot Noir), and American-European hybrid (Traminette) cultivars to seasonal and diurnal variation in field environmental conditions were assessed to propose suitable cultivars for potential use for selection and breeding in grapevines in dry areas. ‘Pinot Noir’ had higher gas exchange parameters, higher water potential, and chlorophyll fluorescence, while ‘Concord’ had the lowest values in all three seasons and experienced moderate stress by the end of each season. The dry conditions in 2005 caused water potential to decline sharply compared to values recorded in 2004 and 2006. ‘Pinot Noir’ and ‘Traminette’ exhibited a mild stress, while ‘Concord’ exhibited a moderate stress in 2006. Photosynthetic rate decreased slightly during the summer with a slight recovery at the beginning of September, and did not differ across the seasons among cultivars. Stomatal conductance followed the same pattern. However, in both 2004 and 2005, stomatal conductance dropped sharply late in the season because of frequently occurring high temperatures that were greater than 32 °C. Maximum morning values of Fv/Fm were within the optimal range. The study suggests the presence of marginal photoinhibition during mid-summer in Concord cultivars, because no full recovery of photochemical efficiency early in the evening was observed. The European cultivar (Pinot Noir) and the American-European hybrid (Traminette) are more adapted to field conditions in Kansas than Concord cultivars. However, high cold-mortality of Pinot Noir due to high sensitivity to cold winter temperature makes the hybrid cultivars the most suitable cultivar for Midwest regions with relatively cold winters.     

Induction of Defense Gene Expression and the Resistance of Date Palm to Fusarium oxysporum f. sp. Albedinis in Response to Alginate Extracted from Bifurcaria bifurcata

In many African countries, the Bayoud is a common disease spread involving the fungus Fusarium oxusporum f. sp. albedinis (Foa). The induction of plant natural defenses through the use of seaweed polysaccharides to help plants against pathogens is currently a biological and ecological approach that is gaining more and more importance. In the present study, we used alginate, a natural polysaccharide extracted from a brown algae Bifurcaria bifurcata, to activate date palm defenses, which involve phenylalanine ammonia-lyase (PAL), a key enzyme of phenylpropanoid metabolism. The results obtained showed that at low concentration (1 g·L⁻¹), alginate stimulated PAL activity in date palm roots 5 times more compared to the negative control (water-treated) after 24 h following treatment and 2.5 times more compared to the laminarin used as a positive stimulator of plant natural defenses (positive control of induction). Using qRT-PCR, the expression of a selection of genes involved in three different levels of defense mechanisms known to be involved in response to biotic stresses were investigated. The results showed that, generally, the PAL gene tested and the genes encoding enzymes involved in early oxidative events (SOD and LOX) were overexpressed in the alginate-treated plants compared to their levels in the positive and negative controls. POD and PR protein genes selected encoding β-(1,3)-glucanases and chitinases in this study did not show any significant difference between treatments; suggesting that other genes encoding POD and PR proteins that were not selected may be involved. After 17 weeks following the inoculation of the plants with the pathogen Foa, treatment with alginate reduced the mortality rate by up to 80% compared to the rate in control plants (non-elicited) and plants pretreated with laminarin, which agrees with the induction of defense gene expression and the stimulation of natural defenses in date palm with alginate after 24 h. These results open promising prospects for the use of alginate in agriculture as an inducer that triggers immunity of plants against telluric pathogens in general and of date palm against Fusarium oxysporum f. sp. albedinis in particular.     

Design,Semisynthesis, Insecticidal and Antibacterial Activities of a Series of Marine-Derived Geodin Derivatives and Their Preliminary Structure–Activity Relationships

To enhance the biological activity of the natural product geodin (1), isolated from the marine-derived fungus Aspergillus sp., a series of new ether derivatives (2–37) was designed and semisynthesized using a high-yielding one-step reaction. In addition, the insecticidal and antibacterial activities of all geodin congeners were evaluated systematically. Most of these derivatives showed better insecticidal activities against Helicoverpa armigera Hübner than 1. In particular, 15 showed potent insecticidal activity with an IC₅₀ value of 89 μM, comparable to the positive control azadirachtin (IC₅₀ = 70 μM). Additionally, 5, 12, 13, 16, 30 and 33 showed strong antibacterial activity against Staphylococcus aureus and Aeromonas salmonicida with MIC values in the range of 1.15–4.93 μM. The preliminary structure–activity relationships indicated that the introduction of halogenated benzyl especially fluorobenzyl, into 1 and substitution of 4-OH could be key factors in increasing the insecticidal and antibacterial activities of geodin.     

Current status and prospects of farming the giant river prawn Macrobrachium rosenbergii (De Man) in Bangladesh

Giant river prawn (Macrobrachium rosenbergii) farming plays an important role in the economy of Bangladesh. Presently, it is cultured in around 50 000 ha area with total annual production of 23 240 t. Traditional extensive prawn farming has been expanding over the last three decades through the introduction and adoption of improved culture systems, such as culture of prawn‐carps, prawn‐shrimp‐fish and prawn‐fish‐rice as concurrent and rotational systems. Efforts for the development of improved techniques on broodstock management, seed production and rearing and grow‐out of prawn have been made over the last decade. The outcomes are low‐cost feed for broodstock, production of post‐larvae in net cages (hapa), all‐male prawn culture, periphyton based prawn‐tilapia culture, C/N based prawn culture, organic prawn farming, prawn‐mola culture and prawn‐carp‐mola polyculture. Despite the development of culture technologies, a number of challenges for sustainable development of prawn farming need to be overcome to realize the potentials of this promising sector. Good aquaculture practises at all levels and application of measures for quality control and food safety would ensure sustainable development of prawn farming in Bangladesh.     

Apparent ileal digestibility of amino acids and the digestible and metabolizable energy content of dry extruded-expelled soybean meal and its effects on growth performance of pigs

We conducted three experiments to determine the apparent ileal digestibility of amino acids (Exp. 1), metabolizable and digestible energy (Exp. 2), and feeding value (Exp. 3) of dry extruded-expelled soybean meal with (DEH) or without (DENH) hulls compared with solvent-extracted soybean meal with hulls removed (SBMNH). Soybeans used to produce DEH were unadulterated prior to extrusion, but those used for DENH were dehulled prior to extrusion. In Exp. 1, six nonlittermate barrows (initially 39 kg) were fitted with ileal T-cannulas and used in a replicated 3 x 3 Latin square design digestion trial. Experimental diets (0.80% total lysine) were cornstarch-based and contained soybean meal from one of the three different sources as the sole source of lysine. Apparent ileal digestibilies of nutrients were similar (P > 0.10) for DEH and DENH. Apparent ileal digestibilies of CP, Lys, Ile, Leu, Arg, Phe, and Val were greater (P < 0.05) for DEH and DENH than for SBMNH. In Exp. 2, six barrows (initially 41 kg) were fed three corn-based diets containing 25% of one of the three soybean meal sources. A fourth diet was fed at the end of the trial containing all ingredients except soybean meal, so that energy values of the soybean meal could be determined by difference. Digestible energy and ME contents were similar (P > 0.10) for DEH and DENH and both had greater (P < 0.05) DE and ME contents than SBMNH. In Exp. 3, pigs (n = 216, initially 10.6 +/- 1.3 kg and 35 +/- 3 d of age) were blocked by weight and allotted to six dietary treatments. Corn-soybean meal-based diets (0.95% digestible lysine and 3.44 kcal/g ME) containing DEH or DENH were compared with similar diets containing SBMNH or solvent-extracted soybean meal with hulls (SBMH). In addition, a diet containing a second expelled soybean meal with hulls (ESBM) was compared with a diet containing SBMH and soy oil. Growth performance of pigs fed diets containing DEH or DENH was not different (P > 0.10) than that of pigs fed corresponding diets containing SMBH or SBMNH. Pigs fed ESBM had lower (P < 0.05) ADG and G/F compared with its corresponding SBMH and soy oil diet. In conclusion, DEH and DENH are more digestible than conventional soybean meal and can be successfully used in swine diets.     

Evaluation of PCR,IEF and ELISA techniques for the detection and identification of potato cyst nematodes from field soil samples in England and Wales

Effective management of potato cyst nematodes (PCNs) requires simple, rapid and accurate identification and quantification of field populations. Soil samples from a survey of 484 fields in potato rotations in England and Wales were used to compare the identification and quantification of PCNs using IEF, PCR, ELISA and bait plant tests. The cyst counts and bait plant test revealed that 64.3% of field samples contained PCNs. Bait plant tests increased the detection rate of PCNs in field samples by 4–6.4%. This means that some infestations are cryptic and would not normally be detected by standard counts. IEF, PCR and ELISA methods distinguished between Globodera rostochiensis and G pallida and were able to register mixed populations; however they were not in full agreement. All methods suggested that G pallida is the dominant species in the field samples tested. The PCR results indicated that 66% of field samples contained pure G pallida, 8% contained pure G rostochiensis and 26% contained mixtures of the two species. Estimates of the relative process times taken per sample in the PCR, IEF and ELISA techniques are given. © 2001 Society of Chemical Industry