Postharvest biocontrol of Alternaria alternata in Chinese winter jujube by Rhodosporidium paludigenum

Aims:  This study was conducted to measure the efficacy of the marine antagonist Rhodosporidium paludigenum in the suppression of postharvest decay of Chinese winter jujube caused by Alternaria alternata and to explore the possible mode of action involved.
Methods and Results:  The efficacy of controlling postharvest diseases by R. paludigenum was examined. Rapid yeast colonization of wounds was observed during the first 48 h at 25°C. The yeast at 1 × 10⁸ cells ml⁻¹ of washed cells suspension provided better control of A. alternata than any other treatment. The concentration of the antagonist had significant effects on biocontrol effectiveness: as the concentration of R. paludigenum was increased, the disease incidence decreased. Meanwhile, R. paludigenum significantly inhibited the natural development of decay and did not damage fruit quality parameters including lightness values, hue angle, firmness, soluble solids, ascorbic acid and titratable acidity in 21 days' storage at 25°C.
Conclusions:  Rhodosporidium paludigenum was effective in controlling postharvest decay of Chinese winter jujube and did not impair fruit quality parameters.
Significance and Impact of the Study:  Rhodosporidium paludigenum can be used as a nonchemical agent in postharvest biological control of Chinese winter jujube.     

Control of postharvest decay on cherry tomatoes by marine yeast Rhodosporidium paludigenum and calcium chloride

Aims: In this study, the potential of calcium chloride (CaCl₂) application to improve the efficacy of the marine antagonist Rhodosporidium paludigenum in controlling postharvest diseases of cherry tomatoes was assessed. Methods and Results: CaCl₂ alone was found not to have any direct influence on the population growth of R. paludigenum in NYDB cultures or in cherry tomato wounds. However, the combined treatments with 1 × 10⁸ cells ml^?1R. paludigenum and CaCl₂ at the concentration from 0·5 to 2% showed high activities to reduce black rot caused by Alternaria alternata in cherry tomato wounds, significantly higher than those of R. paludigenum or CaCl₂ alone. Meanwhile, 0·5% CaCl₂ in combination with 1 × 10⁸ cells ml^?1R. paludigenum greatly inhibited the natural decay of cherry tomatoes in 21 days’ storage at 25°C. Conclusions: The combination of R. paludigenum and CaCl₂ enhances the inhibition of black rot and natural decay of postharvest cherry tomatoes. The results from this study provide a new way to improve the efficiency of R. paludigenum in maintaining the quality of postharvest fruits and vegetables. Significance and Impact of the Study: The marine yeast R. paludigenum combined with CaCl₂ has greatly potential use as an alternative to chemical fungicides in inhibiting postharvest decay on cherry tomatoes.     

STATISTICAL OPTIMIZATION OF A NOVEL LOW-COST MEDIUM BASED ON REGIONAL AGRO-INDUSTRIAL BY-PRODUCTS FOR THE PRODUCTION OF PROTEOLYTIC ENZYMES BY Bacillus cereus

Bacillus sp. are specific producers of peptidase amongst bacteria and peptidase enzymes and are of significant ones due to their multifarious applications. Advances in industrial biotechnology offer potential opportunities for economic utilization of agro-industrial by-products for many biochemical reactions. Due to their rich organic nature, they can serve as an ideal substrate for the production of different value added products like peptidases. In the present work, an attempt was made to optimize different variables by Taguchi methodology for the production of peptidase using agro-industrial by-products hydrolyzed by a Bacillus cereus strain, resulting in brewer's spent grain (BSG) being the optimal organic substrate. Subsequently, operative variables for the BSG were investigated using Taguchi methodology in order to maximize the enzyme production. Additionally, the main medium components were optimized using a mixture design. Finally, the production of peptidase by B. cereus was investigated; also the possible interaction with other proteolytic microbial strains was evaluated. A notorious synergistic effect was observed when B. cereus was inoculated with Pseudomonas sp. These brought a triple benefit, first, opening the possibility to produce technical enzymes at low cost, second, giving greater value to a food industry by-product, and third, reducing the environmental impact caused by the product removal directly into the environment.     

IMPROVED Candida methylica FORMATE DEHYDROGENASE FERMENTATION THROUGH STATISTICAL OPTIMIZATION OF LOW-COST CULTURE MEDIA

NAD⁺-dependent formate dehydrogenase (FDH, EC 1.2.1.2) is of use in the regeneration of NAD(P)H coenzymes, and therefore has strong potential for practical application in chemical and medical industries. A low-cost production of recombinant Escherichia coli (E. coli) containing FDH from Candida methylica (cmFDH) was optimized in molasses-based medium by using response surface methodology (RSM) based on central composite design (CCD). The beet molasses as a sole carbon source, (NH₄)₂HPO₄ as a nitrogen and phosphorus source, KH₂PO₄ as a buffer agent, and Mg₂SO₄ · 7H₂O as a magnesium and sulfur source were used as variables in the medium. The optimum medium composition was found to be 34.694 g L^?1 of reducing sugar (equivalent to molasses solution), 8.536 g L^?1 of (NH₄)₂HPO₄, 3.073 g L^?1 of KH₂PO₄, and 1.707 g L^?1 of Mg₂SO₄ · 7H₂O. Molasses-based culture medium increased the yield of cmFDH about three times compared to LB medium. The currently developed media has the potential to be used in industrial bioprocesses with low-cost production.     

MEDIUM OPTIMIZATION OF ANTIFUNGAL ACTIVITY PRODUCTION BY Bacillus amyloliquefaciens USING STATISTICAL EXPERIMENTAL DESIGN

In order to overproduce biofungicides agents by Bacillus amyloliquefaciens BLB371, a suitable culture medium was optimized using response surface methodology. Plackett–Burman design and central composite design were employed for experimental design and analysis of the results. Peptone, sucrose, and yeast extract were found to significantly influence antifungal activity production and their optimal concentrations were, respectively, 20 g/L, 25 g/L, and 4.5 g/L. The corresponding biofungicide production was 250 AU/mL, corresponding to 56% improvement in antifungal components production over a previously used medium (160 AU/mL). Moreover, our results indicated that a deficiency of the minerals CuSO₄, FeCl₃ · 6H₂O, Na₂MoO₄, KI, ZnSO₄ · 7H₂O, H₃BO₃, and C₆H₈O₇ in the optimized culture medium was not crucial for biofungicides production by Bacillus amyloliquefaciens BLB371, which is interesting from a practical point of view, particularly for low-cost production and use of the biofungicide for the control of agricultural fungal pests.     

STATISTICAL OPTIMIZATION OF GREEN FLUORESCENT PROTEIN PRODUCTION FROM Escherichia coli BL21(DE3)

An optimized cultivation condition is needed to maximize the functional green fluorescent protein (GFP) production. Six process variables (agitation rate, temperature, initial medium pH, concentration of inducer, time of induction, and inoculum density) were screened using the fractional factorial design. Three variables (agitation rate, temperature, and time of induction) exerted significant effects on functional GFP production in E. coli shake flask cultivation and were optimized subsequently using the Box–Behnken design. An agitation rate of 206 rpm at 31°C and induction of the protein expression when the cell density (OD_600nm) reaches 1.04 could enhance the yield of functional GFP production from 0.025 g/L to 0.241 g/L, which is about ninefold higher than the unoptimized conditions. Unoptimized cultivation conditions resulted in protein aggregation and hence reduced the quantity of functional GFP. The model and regression equation based on the shake flask cultivation could be applied to a 2-L bioreactor for maximum functional GFP production.     

我国森林植被的生物量和净生产量

利用森林蓄积量推算森林生物量和净生产量的方法,系统研究了我国森林植被的生物生产力。结果表明,我国森林生物生产力的地理分布规律与世界总趋势一致,但量上有差异,具体表现在：我国森林生物量的平均值小于世界平均水平,而净生产量却显得较高;我国森林的总生物量是９１０２．８７×１０６ｔ,其中,林分为８５９２．１３×１０６ｔ,经济林３２５．７２×１０６ｔ;竹林１８５．０２×１０６ｔ,疏林、灌木林７９０．５４×１０６ｔ;森林和疏林（含灌木林）的总生产力分别是１１７７．３１×１０６ｔ／ａ和４５８．１６×１０６ｔ／ａ。研究结果还显示,用材积推算的生物量（材积源生物量）比用平均生物量计算的结果更符合实际。分析中国森林在中国及全球陆地碳库中的作用发现,与其他区域和世界平均水平相比较,中国森林在中国陆地植被中所起的主导作用较弱,它的生物量不足全球森林总生物量的１％,然而,它在保护中国土壤碳库功能方面起着其他植被类型所无法替代的作用。     

OPTIMIZATION OF FERMENTATION MEDIUM FOR ACETOIN PRODUCTION BY Bacillus subtilis SF4-3 USING STATISTICAL METHODS

To improve the acetoin-producing ability of Bacillus subtilis SF4-3, isolated from “natto,” a Japanese traditional food, the fermentation medium was optimized in shake-flask fermentation by statistically designed methods. Based on results of the single-factor experiment, orthogonal experiment, and Plackett–Burman design, yeast extract, corn steep liquor, and urea were identified as showing significant influence on the acetoin production. Subsequently, the optimum combination of the three factors was investigated by the Box–Behnken design (BBD) of response surface methodology (RSM) in order to further enhance the acetoin production. The maximum acetoin yield of 45.4 g/L was predicted when the concentrations of yeast extract, corn steep liquor, and urea were 8.5 g/L, 14.6 g/L, and 3.8 g/L, respectively. The results were further confirmed in triplicate experiments using the optimized medium (glucose 160 g/L, yeast extract 8.5 g/L, corn steep liquor 14.6 g/L, urea 3.8 g/L, manganese sulfate 0.05 g/L, ferrous sulfate 0.05 g/L), and an acetoin yield of 46.2 g/L was obtained in the validation experiment, which was in agreement with the prediction. After the optimization of medium components, an increase of 36.28% in acetoin production was achieved in comparison to that at the initial medium levels.     

STATISTICAL OPTIMIZATION OF A NOVEL LOW-COST MEDIUM BASED ON REGIONAL AGRO-INDUSTRIAL BY-PRODUCTS FOR THE PRODUCTION OF PROTEOLYTIC ENZYMES BY Bacillus cereus

Bacillus sp. are specific producers of peptidase amongst bacteria and peptidase enzymes and are of significant ones due to their multifarious applications. Advances in industrial biotechnology offer potential opportunities for economic utilization of agro-industrial by-products for many biochemical reactions. Due to their rich organic nature, they can serve as an ideal substrate for the production of different value added products like peptidases. In the present work, an attempt was made to optimize different variables by Taguchi methodology for the production of peptidase using agro-industrial by-products hydrolyzed by a Bacillus cereus strain, resulting in brewer's spent grain (BSG) being the optimal organic substrate. Subsequently, operative variables for the BSG were investigated using Taguchi methodology in order to maximize the enzyme production. Additionally, the main medium components were optimized using a mixture design. Finally, the production of peptidase by B. cereus was investigated; also the possible interaction with other proteolytic microbial strains was evaluated. A notorious synergistic effect was observed when B. cereus was inoculated with Pseudomonas sp. These brought a triple benefit, first, opening the possibility to produce technical enzymes at low cost, second, giving greater value to a food industry by-product, and third, reducing the environmental impact caused by the product removal directly into the environment.     

池鹭繁殖种群数量、活动规律和生物生产量的研究

本文报道池鹭种群数量变动,种群活动,雏鸟生长和生物生产量。种群活动可划分为适到期,运情期,繁殖期,同鸟活动期,集群活动和迁离期。１９９０,１９９１二年繁殖前种群密度分别为１３００只／ｈｍ＾２和１４０１只／ｈｍ＾２,繁殖后增加密度为２０８７只／ｈｍ＾２和２２４８只／ｈｍ＾２。雏鸟体重增长曲线议程为：Ｗ＝２８６／１＋ｅ＾－０．４０９（ｔ－１２．６）１９９０,１９９１年繁殖种群的生物生产量分别为１９７．     

微生物生长稳定性超熵产生判据

运用非平衡热力学超熵产生理论和微生物生长动力学理论,以细胞、营养物和产物为参量,求得了微生物生长的超熵产生及稳定性判据,理论分析与实验计算结果基本一致．     

对采用平均叶龄估算叶年净初级生产量的修正

 根据森林和林木各器官生物量随时间的变化,林冠叶片(叶群体)生长和死亡规律及数学生态学的原理,得出了叶的生物量、年净生产量和平均叶龄之间的关系。一般表达式为P=│lnlx│·B/x(式中Pn为叶年净生产量,lx为特定叶龄的存留率,B为叶生物量,x为平均叶龄)。当林冠叶片达到平衡时,Pn=0.693·B／x;存留率为36.8%时,成为目前用平均叶龄计算叶年净初级生产量的形式。本文证明了后一种特殊形式之所以被人们广泛采用是因为混淆了平均叶龄与叶量周转时间这两个概念;这种方法估算的结果比正常值约高30%。 还用实例讨论了用0—1龄级平均叶龄估算叶净生产量所不可避免的偏大原因,用本文提出的公式对某些叶净生产量的计算数值进行了修正;结果表明,与实际情况比较吻合。     

羊毛生物整理复合酶L86发酵工艺优化研究

采用L9（34）正交试验对L86复合酶生产菌的发酵培养基进行优化、并通过溶氧浓度调节和中期补加蛋白水解液对发酵过程进行调控。结果表明较优的培养基组成为（g/100ml）：黄豆粉4.0、玉米粉1.0、鱼粉0.6、蚕蛹粉0.6、CaCl20.5、NH4Cl1.0、Na2HPO4·2H2O 0.4;通气量控制30h前1:0.5、30-50h1:1.0、50h后1:1.2 v/v/m。在上述条件下摇瓶,酸性蛋白酶酶活10000u/ml、纤维素CMC酶3600u/ml;在0.5m3搅拌罐中扩大中试平均发酵单位酸性蛋白酶5500u/ml、纤维素CMC酶2200u/ml。     

EVIDENCE FOR A HYDRODYNAMIC MECHANISM OF SOUND PRODUCTION BY COURTING MALES OF THE NOTCHTONGUE GOBY,BATHYGOBIUS CURACAO (METZELAAR)

ABSTRACT

Male gobies of the genus Bathygobius are soniferous during courtship. The mechanism by which the sounds are produced is, however, unknown. Early studies on sound production by males of B. soporator suggested that these sounds are hydrodynamic in nature, being produced by the forcible ejection of water through the gill opening. The mechanism of sound production by the closely related species B. curacao was investigated and three lines of evidence are presented which support the hydrodynamic hypothesis. First, similarities between the sounds produced by courting males and by ejecting water through a pipette demonstrated that hydrodynamic forces can readily produce such sounds. Second, the behavioural motor patterns occurring during sound production are consistent with the hypothesis that water is being ejected through the gill openings. Finally, morphological examination revealed an apparent lack of specialised features associated with sound production, effectively eliminating stridulatory and swim bladder mechanisms. These results represent the strongest evidence to date in support of any proposed mechanism of sound production in the gobiids.     

A NEWLY ANTI-Streptococcus suis BACTERIOCIN PRODUCING STRAIN FROM UNWEANED PIGLETS FECAL MATTER: ISOLATION,PRELIMINARY IDENTIFICATION,AND OPTIMIZATION OF MEDIUM COMPOSITION FOR ENHANCED BACTERIOCIN PRODUCTION

A newly isolated anti-Streptococcus suis bacteriocin-producing strain LPL1-5 was obtained from healthy unweaned piglets' fecal matter, and was designated as Lactobacillus pentosus LPL1-5 based on morphology, biochemical properties, and 16S rDNA sequencing analysis. The medium composition for enhanced bacteriocin production by L. pentosus LPL1-5 was optimized by statistical methodology. Yeast extract, K₂HPO₄ · 3H₂O, and MnSO₄ · H₂O were identified as significant components influencing pentocin LPL1-5 production using the Plackett–Burman method. Response surface methodology was applied for further optimization. The concentrations of medium components for enhanced pentocin LPL1-5 production were as follows (g/L): lactose 20.00, tryptone 10.00, beef extract 10.00, yeast extract 14.00, MnSO₄ · H₂O 0.84, K₂HPO₄ · 3H₂O 4.92, triammonium citrate 2.00, Na-acetate 5.00, MgSO₄ · 7H₂O 0.58, Tween 80 1.00. Under the optimized condition, a value of 3154.65 ± 27.93 IU/mL bacteriocin activity was achieved, which was 4.2-fold that of the original medium.