Optimization of Agrobacterium-mediated transformation parameters for Melastomataceae spp. using green fluorescent protein (GFP) as a reporter

Agrobacterium-mediated transformation for both Melastoma malabathricum and Tibouchina semidecandra were optimized using green fluorescent protein (GFP) as a reporter. The binary vector pCAMBIA1304 harboring the modified green fluorescent protein (mgfp) gene driven by the CaMV 35S promoter was used. Parameters optimized were bacterial strain, bacterial concentration, pre-culture period, co-cultivation period, immersion time, acetosyringone concentration and wounding type. Results obtained were based on the percentage of GFP expression which was observed 3 days post-transformation. Agrobacterium tumefaciens strain LBA4404 and EHA105 at concentration 1 × 10⁷ cfu ml⁻¹ (OD_600nm 0.8) showed the highest virulence on M. malabathricum and T. semidecandra, respectively. Four days of pre-culture and 2 days of co-cultivation were optimum for M. malabathricum transformation, while 3 days of pre-culture and co-cultivation for T. semidecandra. Result also showed that 60 min of immersion and addition of 200 μM acetosyringone gave the highest percentage of positive transformants for both M. malabathricum and T. semidecandra. Mild wounding also significantly increased the efficiency of M. malabathricum transformation.     

Effect of exogenous putrescine on post-harvest life of strawberry (Fragaria ananassa Duch.) fruit,cultivar Selva

Effect of different concentrations of putrescine on post-harvest life of strawberry (Fragaria ananassa Duch.) fruit, cultivar Selva at 5 °C was studied. Fruits were immerged in 0.3, 0.5, 1 and 2 mM putrescine as well as distilled water (control) for 5 min, then transferred into the fridge (5 °C) together with untreated fruits (dry treatment). The rate of weight loss, ethylene production, flesh firmness, soluble solids content, titratable acidity and pH of fruits were determined 5, 9 and 13 days after the beginning of storage. Flesh firmness, appearance, color change and taste of fruits were also determined in the same intervals using a taste panel. Storage life of the strawberry fruits was significantly increased by the use of putrescine, so that the untreated and control fruits had 6 and 8 days storage life, respectively, while the immerged fruits in 1 and 2 mM putrescine were still suitable to be exposed in the market 12 and 14 days after the beginning of storage, respectively. No significant weight losses were observed in treated fruits compared to controls and dry treatment at all determination times. Ethylene production was decreased significantly by the use of putrescine. Untreated fruits (dry treatment) had the highest rate of ethylene production and the lowest rate was occurred in 2 mM putrescine treatment at all determination times (5, 9 and 13 days after the beginning of storage). The use of putrescine also prevented the softening of fruit flesh during the storage and kept their firmness, so that, the 2 mM putrescine treatment caused the highest fruit firmness at all determination times. Distilled water treatment (control) had the lowest fruit firmness 5 and 9 days after storage, while this occurred for the dry treatment 13 days after storage. Soluble solids content, pH and titratable acidity of the fruits were not significantly affected by the use of putrescine, but the highest and lowest rate of titratable acidity were related to the 2 mM putrescine and dry treatment, respectively, at the three determination times. Overall, the quality of fruits was improved by the use of 2 mM putrescine in terms of properties evaluated by the taste panel.     

Production of Wild Buffalo (Bubalus arnee) Embryos by Interspecies Somatic Cell Nuclear Transfer Using Domestic Buffalo (Bubalus bubalis) Oocytes

The objective of this study was to explore the possibility of producing wild buffalo embryos by interspecies somatic cell nuclear transfer (iSCNT) through handmade cloning using wild buffalo somatic cells and domestic buffalo (Bubalus bubalis) oocytes. Somatic cells derived from the ear skin of wild buffalo were found to express vimentin but not keratin and cytokeratin‐18, indicating that they were of fibroblast origin. The population doubling time of skin fibroblasts from wild buffalo was significantly (p < 0.05) higher, and the cell proliferation rate was significantly (p < 0.05) lower compared with that of skin fibroblasts from domestic buffalo. Neither the cleavage (92.6 ± 2.0% vs 92.8 ± 2.0%) nor the blastocyst rate (42.4 ± 2.4% vs 38.7 ± 2.8%) was significantly different between the intraspecies cloned embryos produced using skin fibroblasts from domestic buffalo and interspecies cloned embryos produced using skin fibroblasts from wild buffalo. However, the total cell number (TCN) was significantly (p < 0.05) lower (192.0 ± 25.6 vs 345.7 ± 42.2), and the apoptotic index was significantly (p < 0.05) higher (15.1 ± 3.1 vs 8.0 ± 1.4) for interspecies than that for intraspecies cloned embryos. Following vitrification in open‐pulled straws (OPS) and warming, although the cryosurvival rate of both types of cloned embryos, as indicated by their re‐expansion rate, was not significantly different (34.8 ± 1.5% vs 47.8 ± 7.8), the apoptotic index was significantly (p < 0.05) higher for vitrified–warmed interspecies than that for corresponding intraspecies cloned embryos (48.9 ± 7.2 vs 23.9 ± 2.8). The global level of H3K18ac was significantly (p < 0.05) lower in interspecies cloned embryos than that in intraspecies cloned embryos. The expression level of HDAC1, DNMT3a and CASPASE3 was significantly (p < 0.05) higher, that of P53 was significantly (p < 0.05) lower in interspecies than in intraspecies embryos, whereas that of DNMT1 was similar between the two types of embryos. In conclusion, these results demonstrate that wild buffalo embryos can be produced by iSCNT.     

Alternate Two Different Modes of Actions of Green Chemicals to Combat Batrachedra amydraula Meyrick and Cadra cautella Walk on Date Palm Fruit,Egypt

Alternate two different modes of actions of green chemicals： spinosad ＆ methoxyfenozide to combat the lesser date moth, Batrachedra amydraula and the Almond moth, Cadra cautella Walk on dates in Dakhla Oasis, New Valley, Egypt were done in 2010 and 201 I. Based on infestation of the early insect pest B. amydraula, the three-rotation program with basic 1st spray of spinosad 24 SC at 20 mL/100 L showed insignificant difference. Based on minimum effective rotation doses, alternating spinosad 24 SC at 20 mL/100 L, then methoxyfenozide 24 SC at 15 mL/100 L with two-week interval showed zero infestation and 1.8% at the last sample of 2010 and 2011, respectively. Based on infestation of the late insect pest, Cadra spp., tlhe same previous trend of the three-rotation results. Based on minimum effective rotation doses, alternating spinosad 24 SC at 20 mL/100 L, then methoxyfenozid 24 SC at 15 mL/100 L with three-week interval indicated less than 1% and 1% infestation during harvest for 2010 and 201 l, respectively. The last reflected important value when storing the fruit. Both products proved to be working under dry hot high temperatures.     

Denitrification losses from an irrigated sandy-clay loam under a wheat-maize cropping system receiving different fertilizer treatments

Studies were conducted on denitrification in the plough layer of an irrigated sandy-clay loam under a wheat-maize cropping system receiving different fertilizer treatments. The treatments were: N-100 (urea-N at 100kgha^–1year^–1), N-200 (urea-N at 200kgha^–1year^–1), FYM-16 (farmyard manure at 16 tonnes ha^–1year^–1), FYM-32 (farmyard manure at 32 tonnesha^–1year^–1) and the control (unfertilized). Averaged across sampling dates during the wheat season, the denitrification rate as measured by the C₂H₂-inhibition/soil-core incubation method was highest in N-200 (83gNha^–1day^–1), followed by FYM-32 (60gNha^–1day^–1, N-100 (51gNha^–1day^–1), FYM-16 (47gNha^–1day^–1) and the control (33gNha^–1 day^–1). During the maize growing season, average denitrification rate was highest in FYM-32 (525gNha^–1day^–1), followed by FYM-16 (408gNha^–1day^–1), N-200 (372gNha^–1day^–1, N-100 (262gNha^–1day^–1) and the control (203gNha^–1day^–1). Denitrification loss integrated over the whole vegetation period was at a maximum under FYM-32 (13.9kgNha^–1), followed by N-200 (11.8kgNha^–1), FYM-16 (10.6kgNha^–1) and N-100 (8.0kgNha^–1), whereas the minimum was observed for the control (5.8kgNha^–1). Under both crops, denitrification was significantly correlated with water-filled pore space and soil NO₃ ^–-N. The best multiple regression models accounted for 52% and 70% of the variability in denitrification under wheat and maize, respectively. Results indicated that denitrification is not an important N loss mechanism in this well-drained, irrigated sandy-clay loam under a wheat-maize cropping system receiving fertilizer inputs in the range of 100–200kgNha^–1year^–1. Received: 14 January 1997     

寄主龄期、温度对菜蛾盘绒茧蜂与菜蛾啮小蜂种间竞争的影响

在室内调查了分别以2龄、3龄和4龄小菜蛾幼虫作寄主时,及20、26、28和32 ℃下菜蛾盘绒茧蜂和菜蛾啮小蜂间竞争结果.当2龄或3龄小菜蛾幼虫供2种蜂同时产卵寄生时,绒茧蜂的寄生率显著高于啮小蜂的寄生率;分别与其单独寄生时比,前者无显著差异,而后者则显著下降;二种蜂的合计寄生率与任一种蜂单独存在时的相比,以2龄幼虫作寄主时无显著差异,但以3龄幼虫作寄主时则显著提高.当4龄小菜蛾幼虫供2种蜂同时产卵寄生时,绒茧蜂的寄生率与啮小蜂的寄生率之间无显著差异;二种蜂的合计寄生率显著比绒茧蜂单独存在时的高,但与啮小蜂单独存在时的寄生率相比无显著差异.当2种蜂共存时,在所测试的4个温度下,绒茧蜂的寄生率均高于啮小蜂的寄生率,与其单独寄生时的相比,绒茧蜂的寄生率无显著变化,而啮小蜂的寄生率均显著下降.这些结果表明寄主龄期比温度对这两种蜂种间竞争的影响更大.