To study the effect of foliar application of urea fertilizer on the growth and yield of tomato and to find out the optimum concentration of foliar application of urea for maximum growth and yield of tomato, an experiment was conducted with different concentrations of foliar application of urea fertilizer, namely, 2500, 5000, 7500, and 10000 ppm. The experiment was laid out by a randomized complete block design (RCBD) with four replications. Results showed that different yield components and yield of tomato were influenced by the foliar application of different concentrations of urea. The maximum plant height (132.6 cm), number of leaves (30.73), number of green leaves per plant at harvest (21.08), days to first flowering (28.94), number of flower clusters (11.89), number of flowers (75.18), fruit clusters (5.81), fruits per cluster (4.14), and fruits per plant (21.49); length (4.72 cm), diameter (6.58 cm), and weight of individual fruit (151.0 g) were significantly influenced by the 10000 ppm concentration of foliar application of urea fertilizer. The 10000 ppm application gave the highest yield (63.69 t/hm²) with the lowest (28.48 t/hm²) in the control treatment. The yield per plant as well as per hectare increased with increasing concentrations of foliar application of urea fertilizer.

An efficient protein extraction method for two-dimensional gel electrophoresis (2-DE) from plant samples is usually challenging due to the low protein content and high level of interfering compounds. Proteomic analyses of rice (Oryza sativa L.) roots are limited by the lack of an efficient protein extraction method. To establish an effective protocol of protein extraction suitable for 2-DE analysis in rice roots, we evaluated three protein extraction methods (trichloroacetic acid [TCA]/acetone, Mg/NP-40/TCA, and tris-base/acetone). Our results showed that the Mg/NP-40/TCA extraction method had the highest protein yield and is the best resolution of protein separation among the three methods. The TCA/acetone method exhibited clear ?protein? profiles? and ?detected? more ?protein ?spots with the highest intensity in the region of high Mr (above 45 kDa) than the other methods. However, this method was unable to detect proteins with low-Mr (less than 24.0 kDa). The Tris-base/acetone method showed the poorest resolution of protein separation. Our results suggest that the Mg/NP-40/TCA method was the most effective among the three methods and may provide enhanced proteomic information for rice and other crop roots.

A study was undertaken to assess the effect of different storage temperatures on senescence and postharvest performance of isolated flowers of Ranunculus asiaticus L.. The main aim of the work was to develop a cost-effective storage protocol to reduce the postharvest losses and to bring out the transportation of cut flowers of R. asiaticus. The flowers were subjected to two different storage treatments, dry storage and wet storage, and their postharvest performance was compared under laboratory conditions. For this purpose, the buds were harvested at 8:00 AM at loose bud stage (Stage II of flower development). The harvested buds were cut to a uniform size of 15 cm and processed for dry or wet storage. For dry storage, the buds were packed after wrapping them in moistened filter papers and kept at 5oC and 10oC. For wet storage, the buds were held in distilled water in separate glass beakers kept at 5oC and 10oC, respectively. A separate set of buds each for dry and wet storage was kept at room temperature (15±2)oC. After 72 h storage, the buds were kept at room temperature in distilled water. The average life of an individual flower that opened fully was about 4–5 days. The buds kept under wet storage at 5oC and 10oC for 72 h maintained their premature status, while the buds held at room temperature for 72 h generally bloomed. All the buds stored dry maintained their premature status irrespective of storage temperature. Storage of buds for 72 h at 5oC, followed by transferring to distilled water improved the longevity by about 5 to 6 days. Cold storage treatment before transferring to holding solution improved floral diameter, membrane integrity besides maintaining higher fresh and dry mass of flowers, sugar content, soluble proteins, and phenols. Our results suggested that wet and dry storage of premature buds of R. asiaticus for 72 h at 5oC, followed by placing them in distilled water, improved the cut flower longevity and can be used as effective postharvest storage treatments for this beautiful cut flower.

Glandless upland cotton has an important economic value. Embryogenic calli and regenerated plants were obtained from the hypocotyl explants of glandless upland cotton seedlings, cultivar Jisheng1. The results indicated that somatic embryogenesis was significantly influenced by the types of auxin and cytokinin. 2, 4-D was advantageous to induce cotton callus, but embryogenic callus could not be obtained on the 2, 4-D medium. Embryogenic calli were also not obtained on the MSB sold medium with the combination of IBA and BA. However, embryogenic calli were induced when the hypocotyl explants were cultured on the IBA and KT medium. More than 31% of the hypocotyl segments produced embryogenic calli when the MSB medium was supplemented with 1.0 mg·L^-1 IBA and 0.5 mg·L^-1 KT. Embryogenic calli with somatic embryos could be observed within three months. Somatic embryo germination and maturation occurred on the hormone-free MSB medium with 1.0 g·L^-1 Gln and 0.5 g·L^-1 Asn. A number of regenerated plants could be obtained in six months. In the present study, a simple and efficient system was established to induce a number of embryogenic calli and regenerate plantlets from hypocotyl explants.

The occurrence of fruit shrink disease in Ziziphus jujuba Mill. ‘Linyilizao’ was investigated from 2006 to 2009 in Cangxian of Hebei Province, Taiyuan, and Taigu of Shanxi Province, and the pathogens were isolated from the diseased fruits sampled from both Cangxian and Taiyuan in 2006 and 2007. The pathogens in leaf, fruit, and shoot collected from Cangxian were isolated from June to October in 2008 at 30-day intervals. The incidence of the disease varied significantly with location and year, and the highest incidence was observed in Cangxian during 2006. Three kinds of key pathogens were isolated from the fruits from July to September, i.e., Alternaria alternata Keissler (mainly in July), Phoma destructiva Plowr. (mainly in September), and Fusicoccum sp., of which A. alternata was the most dominant. In leaves, both A. alternata (much more dominant) and P. destructiva could be isolated throughout June to October, but no Fusicoccum sp. was found. At the white-ripening stage of fruit, the isolation rate of A. alternata was the highest in leaves (36.67%), followed by extension shoot (6.67%) and fruit (1.11%) while that of P. destructiva was the highest in extension shoot (31.11%), followed by fruit (13.33%) and leaf (12.22%), with Fusicoccum sp. isolated only in extension shoot (2.22%). The frequency of isolating two or three kinds of above three pathogens together was very low, with the highest rate (19.30%) found in A. alternata and P. destructiva from leaves in July.

Analysis of genetic interactions between rice and its pathogenic fungi Magnaporthe oryzae and Rhizoctonia solani should lead to a better understanding of molecular mechanisms of host resistance, and the improvement of strategies to manage rice blast and sheath blight diseases. Currently, dozens of rice resistance (R) genes against specific races of the blast fungus have been described. Among them, ten were molecularly characterized and some were widely used for breeding for genetic resistance. The Pi-ta gene was one of the best characterized rice R genes. Following the elucidation of its molecular structure, interaction, distribution, and evolution, user friendly DNA markers were developed from portions of the cloned genes to facilitate the incorporations of the Pi-ta mediated resistance into improved rice varieties using marker assisted selection (MAS). However, rice blast is still a major threat for stable rice production because of race change mutations occurring in rice fields, which often overcome added resistance based on single R genes, and these virulent races of M. oryzae pose a continued challenge for blast control. For sheath blight, progress has been made on the exploration of novel sources of resistance from wild rice relatives and indica rice cultivars. A major quantitative trait locus (QTL), named qSB9-2, was recently verified in several mapping populations with different phenotyping methods, including greenhouse methods. The ability to identify qSB9-2 using greenhouse methods should accelerate the efforts on the qSB9-2 fine mapping and positional cloning.

Two approaches of genetic analysis of quantitative traits were compared with a case study on soybean. One approach was the segregation analysis developed by Gai et al. (2003), which utilized information from individuals of one or multiple segregation populations as well as that from parents based on the principles of the major-gene plus polygene inheritance model, mixture distribution, joint maximum-likelihood function, IECM (Iterated Expectation and Conditional Maximization) algorithm, and Akaike’s information criterion and goodness of fit tests. Another approach was quantitative trait locus (QTL) mapping with molecular markers. A recombinant inbred line (RIL) population with 201 families derived from (Kefeng No.1x1138-2) F_2:7:10 along with their parents were tested in a randomized block design experiment. The 171 RFLP, 60 SSR, and 79 AFLP molecular markers were used to mark the 201 families. The data of nine traits, i.e., number of days to flowering, number of days to maturity, plant height, number of nodes on main stem, number of pods per node, 100-seed weight, protein content, oil content, and plot yield, were analyzed with the segregation analysis procedure of RIL population with parents (Gai et al., 2003; Zhang and Gai, 2000; Zhang et al., 2001) to detect their genetic system, and those along with the molecular marker data were analyzed with WinQTL Cartographer (Basten et al., 1999; Zeng, 1993, 1994) to detect their QTL system. The results showed that both procedures could detect the main major genes or QTLs, and therefore, could be used as a mutual check and supplement. From the results that most of the traits were mainly controlled by three or four QTLs, it was impressed that the segregation analysis procedure of four major-gene plus polygene mixed inheritance model should be developed to fit the requirements. The results also showed that the QTLs of the involved traits concentrated on several linkage groups, such as C2, B1, F1, M, and N. Finally, the results showed that the experimental sample was not necessarily coincident with the theoretical population according to equality test, symmetry test, and representation test, and therefore, the sample should be checked, tested and then adjusted to fit the theoretical requirements through deleting the extra-biased families and markers.

A multiplex PCR assay system was developed for the detection of Clavibacter michiganensis subsp. michiganensis (Cmm), which combined two tests in one reaction mixture. Cmm-specific primers PSA-4/PSA-R and Solanum lycopersicum–specific primers NS-7-F/NS-8-R (internal PCR control primer) were combined in one PCR reaction mixture with Cmm and plant DNA as template. The primer sets could amplify the target product successfully. Different combinations and concentrations of primers and annealing temperatures were tested, respectively. The detection level of the optimized multiplex PCR assay was up to 5×10² cfu·mL^-1. To verify the applicability of this system, it was employed to detect Cmm in tomato seeds and plantlet samples. Seeds mixed with Cmm and diseased plantlets were detected successfully. The multiplex PCR system will avoid false-negative results and provide a reliable method for the detection of Cmm.

The effects of incorporation SA into KNO₃ priming solution on the germination and emergence of eggplant seeds at 15°C were investigated. Seeds were primed into 3% KNO₃ containing 0.05, 0.1, 0.5, or 1 mM salicylic acid (SA) for 6 days, respectively. After the priming, seeds were either immediately used for germination and emergence test at 15°C or stored at 4°C for 1 month and then for the germination test. The primed eggplant seeds in general improved the final germination percentage (FGP), germination rate (G₅₀), and germination synchrony (E_10–90) at 15°C compared with non-priming seeds. Priming seeds in 3% KNO₃ solution supplement with 0.1 mM SA resulted in the best priming effect compared with other priming treatments and non-priming treatment, and all of the beneficial priming effects were still retained after stored at 4°C for 30 days. These results indicated that priming seeds in 3% KNO₃ solution containing 0.1 mM SA could be used as an effective method to improve low-temperature performance of eggplant seeds and subsequent seedling growth.

A novel strain of Bacillus thuringiensis Bt11, isolated from soil samples in China, was classified and characterized in terms of its crystal proteins, cry genes content. The Bt11 strain showed high toxicity against Spodoptera exigua and Helicoverpa armigera neonates. Bt11 strain shares morphological and biochemical characteristics with the previously described Bacillus thuringiensis subsp. kurstaki. SDS-polyacrylamide gel electrophoresis revealed that crystals were composed of several polypeptides ranging from 20 to 130 kDa, of which the 35, 80, and 130 kDa proteins were the major components. PCR-RFLP with total DNA from strain Bt11 and specific primers for cry1, cry2, cry3, cry4/10, cry7, cry8, cry9, and cry11 genes revealed that cry1Aa, cry1Ab, cry1Ia, and cry9Ea genes were present.

Opium poppy (BoldItalic L.), an important medicinal plant, produces several opiate alkaloids including morphine, thebaine, codeine, papaverine and noscapine. Polysaccharides, such as sodium alginate, have been used in depolymerized form as wonderful promoters of plant growth. The present study has revealed that application of alginate oligosaccharides (AO), obtained from sodium alginate irradiated by Co-60 gamma rays, significantly enhances certain physiological/biochemical parameters as well as the overall growth of opium poppy. The highest dose applied was the most effective in increasing the morphine and codeine contents as well as the overall yield of crude opium per plant.

The postharvest physiology of strawberry was investigated to study the effects of ozone on weight loss rate, ascorbic acid, respiration rate, peroxdase (POD) activity, catalase (CAT) activity, and malondialdehyde (MDA) content during cold storage. The strawberries were treated with ozone of 0 ppm (control), 2 ppm, 4 ppm, and 8 ppm, respectively. The results indicated that the treatment of 4 ppm ozone could inhibited the decrease of ascorbic acid, POD activity, and CAT activity, and reduced weight loss rate and MDA content. The treatment delayed the senescence of strawberry, with a significantly lower respiration rate. Thus, the best concentration of ozone was 4 ppm, and ozone treatment could be a good candidate for maintaining postharvest quality of strawberry and provide a longer storage life.

In this paper, seed morphology and effects of pre-sowing treatments were studied. Matured seeds of the species were collected from healthy trees in the National Botanical Garden, Bangladesh, and treated with five pre-sowing treatments. The average length, breadth and thickness were found to be 0.56±0.03 cm, 0.44±0.009 cm and 0.26±0.008 cm in Albizia richardiana King and Prain, and 1.32±0.02 cm, 0.55±0.04 cm and 0.11±0.002 cm in Lagerstroemia speciosa L., respectively. Germination was carried out in polybags with a mixture of topsoil, coconut husk compost, coarse sand and fine sand in a ratio of 3∶4∶1∶1. Results revealed that the germination rates of seeds in different pre-sowing treatments were significantly increased compared to those in cold-water treatment in both species. The highest germination rate was found to be 96% in hot-water treatment followed by 87%, 83% and 49% in treatments with scarification, H₂SO₄ and control in A. richardiana, respectively. However, the highest germination rate (79%) was found in H₂SO₄ treatment followed by 64%, 62% and 25% in treatments with hot water, scarification and control in L. speciosa, respectively. The lowest germination (35% in A. richardiana and 20% in L. speciosa) was found in cold-water treatment in both cases. Analysis of variance (ANOVA) showed the significant difference among the treatments for both species. It is concluded that hot-water treatment is recommended for seed germination of both species in rural Bangladesh.

Field experiments were conducted with mustard (Brassica campestris cv. B₉) consecutively for three years (2005, 2006 and 2007) in the Crop Research and Seed Multiplication Farm of Burdwan University, Burdwan, West Bengal, India. In the first year, varietal screening of mustard under recommended dose of chemical fertilizer (100∶50∶50) were performed with seven mustard varieties during November 2005 to February 2006. In the second year, two experiments were conducted to study the effects of six different combined doses of chemical fertilizer and biofertilizer and six different levels of growth retardant cycocel. In the third year, six different levels of compost along with the best screened dose of growth retardant (CCC-300 ppm) and the best combined dose of biofertilizer and chemical fertilizer (3/4thchemical fertilizer:1/4th biofertilizer) from the previous year were applied to study the effects on agronomic traits and growth attributes of mustard, and the field data were analyzed statistically. The variety B₉ recorded a significant higher seed yield along with other yield contributing factors, which was found to be superior to other varieties under old alluvial soil of Burdwan, West Bengal, India, in 2005. In 2006 and 2007, seed yield was found to be the best for CCC-300 ppm treatment and the best combined dose of biofertilizer and chemical fertilizer was found to be 3/4th chemical fertilizer:1/4th biofertilizer. In 2007, the best yield was given by the treatment of 7.5 t·hm^−2 compost along with the best dose of growth retardant cycocel and the best combined dose of biofertilizer and chemical fertilizer.

Half diallel analysis of eight parents was carried out to identify the high heterotic crosses and their relationship in terms of general and specific combining ability (GCA & SCA) in Brassica juncea L. Czern and Coss at IARI, New Delhi, during 2007–2008 and 2008–2009. The relative heterosis and heterobeltiosis were observed to be the highest with respect to seed yield per 100 siliquae and days from sowing to 50% flowering in case of cross IC-199715 × IC-199714, EC-289602 × Prakash in the number of primary branches per plant and harvest index, Agra Local × Pusa Bahar in the length of main axis, Poorbijaya × Agra Local in the number of siliquae on main axis and EC-289602 × Pusa Bahar in the biologic yield per plant and seed yield per plant. Different cross combinations exhibited the maximum value of better and mid-parent heterosis for the remaining traits, viz., days to maturity, number of secondary branches per plant, plant height and 1000-seed weight. GCA and SCA variances were significant in all characters. The variance of GCA (s²g) was observed to be higher from sowing to 50% flowering and maturity in plant height and 1000-seed weight, whereas the variance of SCA (s²s) was higher in seed yield and other remaining parameters.

The fungal pathogen Botrytis cinerea Pers. causes severe rotting on tomato fruits during storage and shelf life. As a biological control agent, endophytic bacterium was regarded as an effective alternative to chemical control. Out of 238 endophytic bacterial isolates, three strains (EB-15, EB-28, and EB-122) isolated from Lycopersicum esculentum Mill., Speranskia tuberculata (Bge.) Baill, and Dictamnus dasycarpus Turcz. respectively were found to be strongly antagonistic to the pathogen in vitro and were selected for further in vivo tests. One endophytic bacterium strain, encoded EB-28, was selected from the three in vivo tested isolates. The inhibitive rate of EB-28 reached 71.1% in vitro and 52.4% in vivo. EB-28 was identified as Bacillus subtilis according to its morphological, physiological, and biochemical characteristics and 16S rDNA sequence analysis.

Sweet cherry (Prunus avium L.) is a kind of fruit with short postharvest life. Postharvest treatment of 1-methylcyclopropene (1-MCP: 1.0, 1.5 and 2.0 μL/L) was applied to cherry fruits, and its effects on postharvest fruit quality during 60-day-storage at cold temperature was investigated. Sweet cherry fruits were harvested at commercial fruit maturity and exposed to 1-MCP at 20±1°C for 24 h. Following 1-MCP treatments, the fruits were stored for 60 d at 90±5% RH and, 0±1°C, in three concentrations of 1.0, 1.5, and 2.0 μL/L 1-MCP, of which 1.5 μL/L 1-MCP had the most positive effect on fruit quality. Postharvest treatments of 1-MCP significantly reduced endogenous ethylene production, malondialdehyde (MDA) content, and polyphenol oxidase (PPO) activity in cherry fruits 60 d after cold storage when compared to untreated fruits. 1-MCP treatment was associated with lower respiration rate and relative electric conductivity (REC) and maintained cell membrane integrity. The results indicated that the treatment 1-MCP was effective in inhibition of the declining of POD activity and CAT activity. In conclusion, 1-MCP treatment could be a good candidate for maintaining postharvest quality of cherry, and 1-MCP could prolong the storage life of sweet cherry.

Zn is an essential mineral nutrient for plant growth and development. Its effect on crop yield and quality has not been well documented. The objective of this study was to investigate the effects of foliar application of zinc fertilizers on yield and quality of millet (Pennisetum glaucum). Six levels of ZnSO₄·7H₂O (0, 0.75, 1.13, 1.50, 2.25, 3.00 kg/hm²) were applied to two millet cultivars (Jigu 20 and Jiyou 2) to obtain 12 treatments. Zinc fertilization increased millet yield and improved quality when applied at a proper rate in both cultivars. The yield was the highest in Jigu 20 when applied at 1.50 kg/hm², and in Jiyou 2 when treated with 1.50 kg/hm². Zn application at 1.5 kg/hm² increased protein content by 11.13% for Jigu 20 and 10.53% for Jiyou 2. The Zn application at all rates increased lysine acid and soluble sugar content in the grain in both cultivars. The results of this study suggest that foliar Zn application increases yield and also improves grain quality when applied at 1.50 to 2.25 kg/hm² for soils with low zinc content.

The objective of this research was to develop a detection method for Eperythrozoon wenyoni infection using polymerase chain reaction (PCR) assay technique. A pair of primers was designed and synthesized according to the conservative sequence 16S rRNA. The PCR assay was performed with the primers. A 985-bp fragment was amplified by using PCR. The amplified fragments with the expected size were identified by EcoR I restriction digestion. The crossing-reaction, specific-reaction and duplicate-reaction indicated that the PCR method is a specific, sensitive, fast and effective method for diagnosing E. wenyoni infection at group level.

A systematic and comparative study on the frequency and spectrum of chlorophyll mutations induced by ethylmethane sulphonate (EMS) - an alkylating agent, hydrazine hydrate (HZ) – a base analogue and sodium azide (SA) – a respiratory inhibitor, was carried out in two mungbean varieties, namely, PDM-11 and NM-1. A wide spectrum of chlorophyll mutants was obtained in the M₂ generation. All these chlorophyll-deficient mutants were lethal except maculata, viridis and virescent. Chlorina followed by xantha types were predominant in both the varieties. EMS treatments induced the highest frequency of chlorophyll mutations followed by HZ and SA. The frequency of chlorophyll mutations was dose-dependent and increased with the mutagen concentration. Based on effectiveness in both varieties, the order of mutagens was HZ>SA>EMS. Two criteria viz., pollen sterility (Mp/S) and seedling injury (Mp/I) were taken into consideration to determine the efficiency of the mutagens. EMS was found to be the most efficient mutagen followed by HZ and SA. Moderate concentrations of the mutagens were the most effective and efficient in inducing mutations.

Phytosociological parameters, soil and temperature conditions, importance values of species, life form, leaf size, and biomass were investigated at the village Tabai during autumn 2006. There was very little difference in air and soil temperature due to similar elevation. There was a great difference in biomass production for various stands. The original vegetation structure has been altered due to deforestation and overgrazing. There is a need for restoration of the habitat.

In the long run, whether the use of chemical fertilizers could be reduced and soil fertility could be maintained through rice–fish coculture is less well known. At the pilot site of the rice–fish coculture system, which is one of the five “globally important agricultural heritage systems” (GIAHS), we conducted a 4-year study to compare fertilizer use, rice yield, and soil fertility in rice–fish coculture and rice monoculture. Based on the survey data from 21 villages, rice yield did not differ between rice monoculture and rice–fish coculture, but less chemical fertilizers were used in rice–fish coculture than in rice monoculture. Survey data from 145 farms also showed that rice–fish coculture farms with high input of feed for fish used less chemical fertilizers for rice production than farms with low input of feed for fish. In the 4-year field experiment, although less fertilizer was used in rice–fish coculture, rice yield, soil organic matter, soil total nitrogen, and soluble phosphorus did not differ between rice–fish coculture and rice monoculture. Our results suggest that rice–fish coculture can reduce chemical fertilizers application, enhance land productivity, and maintain soil fertility. Our results also suggest that rice–fish coculture could reduce the risk of non-point source pollution by reducing the input of chemical fertilizers.

Radishes (Raphanus sativus L.) were grown in plastic pots in a screenhouse to investigate the influences of nitrogen fertilizer application rates (NFAR) on yield, nitrate content, nitrate reductase activity (NR), nutrition quality, and nitrogen recovery efficiency (NRE) at commercial mature stage. Five N-rate treatments, 0.644, 0.819, 0.995, 1.170, and 1.346 g?pot^-1, were set up in the screenhouse pot experiments, and nitrogen fertilizer (unlabeled N and ¹⁵N-labeled fertilizer) was applied as basal dressing and topdressing, respectively. The results indicated that the fresh and dry weight yields of radish increased with the increase of NFAR at the range of 0.099 to 0.180 g N?kg^-1 soil, decreased at 0.207 g N?kg^-1 soil, and accordingly there was a significant quadratic relationship between the fresh and dry weight yields of radish and the NFAR. At the high addition of urea-N fertilizer, the nitrate content accumulated in the fleshy roots and leaves due to the decline in NR activity. From 0.644 to 0.819 g N?pot^-1 NR increased most rapidly, the highest NR activity occurred at 0.819 g N?pot^-1, and the lowest NR activity happened at 1.346 g N?pot^-1. Soluble sugar and ascorbic acid initially increased to the highest value and then decreased, and, contrarily, crude fiber rapidly decreased with the increase of NFAR. Total N uptake (TNU), N derived from fertilizer (N_dff), and N derived from soil (N_dfs) in radish increased, except that N_dfs relatively and slightly decreased at the rate of 0.207 g N?kg^-1soil. The ratio of N_dff to TNU increased, but the ratio of N_dfs to TNU as well as NRE of N fertilizer decreased with the increase of NFAR. Therefore, the appropriate NFAR should be preferably recommended for improving the yields and nutrition qualities of radish and NRE of N fertilizer.

Site-directed mutagenesis is used extensively for probing gene function. In this paper we describe an improved megaprimer method to the site-directed mutagenesis of phytase from Aspergillus niger, which allowed the mutations to be performed more efficiently in less time than other traditional methods. Three rounds of PCR and two pairs of primers were required in this method, and additionally, the restriction enzyme Dpn I was used for the elimination of template instead of the gel purification in this process. The entire procedure was performed in one tube. Moreover, this method was easier for obtaining large mutant genes than other methods. We successfully carried out the site-directed mutagenesis of phytase by adopting this method.

Some changes in color and morphology of Anoplophora glabripennis larvae were observed when infected with Metarhizium (Metsch.) Sorokin MS01. Paraffin sections show the spores of MS01 attached to the cuticle of A. glabripennis larvae at 24 h post inoculation (hpi). At 36 hpi, the conidium germinated and penetrated through the cuticle. After 48 h of inoculation, the endocuticle was disintegrated and the germ tubes passed through the epithelial layers into the hemocoel and multiplied, and then invaded the tissues near the penetration point, such as the fat body, muscle, tracheal tissue, malpighian tubule, alimentary canal, etc. The infected larvae were dead and most tissues and organs were infected and disintegrated with the multiplication of the mycelium for 3 days. The hyphae emerged through the cuticle and formed a hyphal layer on the surface of the dead insect and the conidium emerged five days after inoculation.

The purpose of this study was to assess the synergistic effects of exogenously applied proline and glycinebetaine (betaine) in antioxidant defense and methylglyoxal (MG) detoxification system in mung bean seedlings subjected to salt stress (200 mmol·L^-1 NaCl, 48 h). Seven-day-old mung bean seedlings were exposed to salt stress after pre-treatment with proline or betaine. Salt stress caused a sharp increase in reduced glutathione (GSH) and oxidized glutathione (GSSG) content in leaves, while the GSH/GSSG ratio and ascorbate (AsA) content decreased significantly. The glutathione reductase (GR), glutathione peroxidase (GPX), glutathione S-transferase (GST) and glyoxalase II (Gly II) activities were increased in response to salt stress, while the monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), catalase (CAT) and glyoxalase I (Gly I) activities sharply decreased with an associated increase in hydrogen peroxide (H₂O₂) and lipid peroxidation level (MDA). Proline or betaine pre-treatment had little influence on non-enzymatic and enzymatic components as compared to those of the untreated control. However, proline or betaine pre-treated salt-stressed seedlings showed an increase in AsA, GSH content, GSH/GSSG ratio and maintained higher activities of APX, DHAR, GR, GST, GPX, CAT, Gly I and Gly II involved in ROS and MG detoxification system as compared to those of the untreated control and mostly also salt-stressed plants with a simultaneous decrease in GSSG content, H₂O₂ and MDA level. These results together with our previous results suggest that coordinate induction of antioxidant defense and glyoxalase system by proline and betaine rendered the plants tolerant to salinity-induced oxidative stress in a synergistic fashion.

This article discusses a range of studies on starch changes in tobacco leaves during flue-curing. Important effects resulting from using different flue-curing equipment and techniques on starch content are presented. It concludes that future researches should be focused on the structure and physico-chemical properties of tobacco starch during the curing process.

Annual monitoring of the population dynamics of the oriental fruit fly Bactrocera dorsalis (Hendel) (Diptera: Tephritidae) in Xishuangbanna, southern Yunnan, was conducted by using methyl eugenol-baited traps in 1997, 2000, and 2003, and factors including temperature, rainfall, and host species with respect to the population fluctuation were analyzed systematically. The results showed that the fruit fly was present all year round in Xishuangbanna. Its population remained at a lower level from November to February, and increased from March until it reached a peak in June or July, depending on the rainfall that year. Afterward, the fly population declined remarkably until October. Temperature, rainfall, and host fruits were major factors comprehensively influencing the population fluctuation. The monthly mean temperature was in a range of temperatures suitable for development and reproduction of the fly. But the monthly mean minimum temperature from December to February was lower than the suitable temperature range, which was suggested a possible reason for the lower populations in winter months. Rainfall was another essential factor influencing the population fluctuations. The population was depressed when the monthly rainfall amount was lower than 50 mm, but increased when rainfall ranged from 100 mm to 200 mm. When the monthly rainfall amount was higher than 250 mm, the fruit fly population was reduced remarkably. The heavy rain in July and August explained the decreasing population. Mango, orange, pear, longan, and peach were found to be the main host species of the fly in Xishuangbanna. Among them, mango and longan were most preferred by the fly. Therefore, the planted areas, fruiting period, and production exerted essential effects on the fly population fluctuations, which were regarded as another major factor influencing the fly population in that area. Briefly, temperature, monthly rainfall, and the host species, through the way of their functions, the influence strength, as well as the period that they occurred comprehensively impacted the population fluctuations of the fruit fly in Xishuangbanna.

The effects of soil drought on soluble protein content and protective enzyme system of cotton leaves at different positions were studied in the transgenic cotton cultivar, Lumianyan28. The results indicated that the soluble protein content in main stem leaves and in middle-fruit branch leaves under drought treatment were higher than that of CK, the normal soil water management treatment. Lower fruit branch leaves under drought treatment had higher protein and enzyme levels than CK treatments. From June 25 to July 22, the activity of superoxide dismutase (SOD) in main stem leaves under drought treatment was lower than that of CK. Other time periods had varying results. The peroxidase (POD) activity in main stem leaves and in fruit branch leaves was lower at early stage and then higher at late stage, showing a trend of descending first, and then ascending obviously under the drought condition. The catalase (CAT) activity showed an increase-decrease trend, higher in main stem leaves under drought treatment than that of CK. However, it was opposite in the lower fruit branch leaves, and there were no significant differences between the two CAT treatments in the middle branch leaves. It is suggested that the soluble protein and cellular protection enzymes, such as superoxide dismutase, peroxidase activity, and catalase in main stem leaves and fruit branching leaves play important physiological functions in the early growth stage under drought stress.

The essential oil from the leaves of yacon grown in China was isolated by hydrodistillation and distillation-extraction. Chemical constituents of the essential oil were separated and identified by means of gas chromatography/mass spectrometry (GC/MS) for the first time, and the relative content of each constituent was determined by area normalization. Twenty-one chemical constituents were identified, and their amounts accounted for 96.2% of the total composition. The main components of the essential oil were β-phellandrene (26.3%), β-cubebene (17.7%), β-caryophyllene (14.0%) and β-bourbonene (10.2%). Therefore, in the volatile oil from the leaves of yacon, sesquiterpenes are major compounds, accounting for 52.2%.