Saccharification of cellulose by dry pyrolysis

Pyrolysis of cellulose was studied for the purpose of practical production of 1,6-anhydro-β-D-glucopyranoside (levoglucosan, LG). To minimize secondary degradation of levoglucosan, two methods were examined: (1) conductive heating by glass bottle, and (2) radiation heating from the surface by CO₂ laser beam, both under vacuum and in a nitrogen atmosphere. Glass-bottle pyrolysis under vacuum gave levoglucosan yield of 50%–55% in the optimum temperature range of 350°–410°C, where placing the cold trap in the vicinity of heated area was effective in improving the yield. In contrast, glass-bottle pyrolysis under nitrogen gave low yields of 17%–20%, probably due to slower diffusion of pyrolysis product from hot region. The CO₂ laser pyrolysis under vacuum gave the product as aerosol (white smoke), causing difficulty in recovery of the product, and the maximum yield was 5%–17%. In this case the treatment under nitrogen flow was effective for recovery of aerosol, and the maximum yield reached approximately 25%.     

Exposure to probiotics and β-1,3/1,6-glucans in larviculture modifies the immune response of Penaeus vannamei juveniles and both the survival to White Spot Syndrome Virus challenge and pond culture

The effect of the inclusion of probiotics (Vibrio alginolyticus) and β-1,3/1,6-glucans in Penaeus vannamei larviculture was evaluated by measuring the immune response and survival of shrimp juveniles subjected to white spot syndrome virus (WSSV) challenge tests (per os) and pond culture. Treatments were designed to contrast the probiotic factor (inclusion vs non-inclusion) and β-1,3/1,6-glucans supplementation in various larvae stages; starting from early stage (Zoea II), to middle stage (PL 12), late stage (15 days pre-challenge) or non-supplemented. In larviculture, the highest survival was obtained in animals treated with β-1,3/1,6-glucans from Zoea II. The use of probiotics enhanced survival during the first 0-52 h post-WSSV challenge period. During 56-156 h post-WSSV challenge period, interactions were observed between β-1,3/1,6-glucans × time and β-1,3/1,6-glucans × time × probiotics. Significant differences in final survival rates between treatments were not observed. In the second WSSV challenge, immune parameters were analysed. Significant interaction between probiotics and β-1,3/1,6-glucans was observed for plasmatic protein (PP) concentration, super oxide anion (O₂⁻) generation, antibacterial activity (AA), and total haemocyte count (THC). The use of probiotics in larviculture had a negative effect on the PP, but increased the AA and THC, while β-1,3/1,6-glucans had a negative effect on the O₂⁻ generation. The most relevant results were obtained from treatments T2 (probiotics in larviculture, β-1,3/1,6-glucans from Z II) and T4 (probiotics in larviculture, β-1,3/1,6-glucans 15 days before challenge). Treatment T2 presented the highest survival rate in larviculture. After WSSV infection, the animals of this treatment displayed resistance to the virus, a strong AA and increase of THC. Treatment T4 increased the amount of PP, increased the O₂⁻ generation and THC. Histological analysis showed that the animals of treatment T2 and T4 were able to limit the spread of the virus during the first hour after challenge with WSSV. The survivors from treatments T2 and T4 had a high THC, accompanied by a lack of white spot disease (WSD) injuries. A bioassay was carried out under farm conditions during the warm-rainy season using larvae from treatments T2 and T4. The animals were stocked at 18 animals/m² in earth ponds of 0.20 ha (three ponds/larvae kind). WSD outbreak was not detected, and the survival was significantly higher in ponds stocked with larvae from treatment T4 (70 ± 3%) than in ponds stocked with larvae from treatment T2 (49 ± 9%).     

Distribution characteristics and diversities of cbb and coxL genes in paddy soil profiles from southern China

The Calvin-Benson-Bassham cycle and Wood-Ljungdahl pathway are two well-known ones in the six carbon sequestration pathways, but the current knowledge of their occurrence in different layers of agricultural soil profiles is poor. In this study, the diversities of three genes encoding ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO), i.e., genes encoding the green-like (cbbLG) and red-like (cbbLR) forms of RubisCO I and encoding RubisCO II (cbbM), and the gene encoding carbon monoxide dehydrogenase large subunit (coxL) from five paddy soils in southern China were investigated by real-time quantitative polymerase chain reaction, restriction fragment length polymorphism (RFLP) analysis, and clone library. The abundances of the four genes ranged from 10⁷ to 10⁹ copies g^-1 soil, and the cbbLR gene outnumbered the other three genes in all soil samples, suggesting important roles they play in carbon dioxide (CO₂) fixation. In addition, it was found that the copy numbers of cbbLR and cbbLG decreased with increasing soil depth, while the copy numbers of cbbM and coxL decreased in the shallow depths but increased with increasing soil depth. The results of RFLP showed a larger Shannon index (H) in the deeper soil layers among the four gene clone libraries, indicating that the community diversity in these soil layers was greater. The cbbLG gene had relatively low diversity (at genus level), and most of the sequences were classified as Sideroxydans and Thiobacillus. In contrast, the highly diverse groups were found in the other three gene clone libraries (cbbLR, cbbM, and coxL), most of which were distantly related to known sequences, even forming separate clusters. In summary, this study provides a new insight into CO₂ fixers along agricultural soil profiles by comparing four bacterial genes.     

β-1,3/1,6-葡聚糖在畜禽日粮中的应用效果

β-1，3/1，6-葡聚糖作为一种潜在的免疫调节佐剂，普遍存在于真菌、酵母及海藻中，主要由以β-1，3键构成的线性骨架为主链，以及以高度分支的β-1，6键为侧链组成，其特殊的键结方式和分子内氢键使其形成螺旋形的分子结构，这种独特的构型很容易被免疫系统识别，从而发挥免疫功能。β-1，3/1，6-葡聚糖是第一个被发现的具有免疫活性的葡聚糖，其作为一种生物性免疫调节佐剂，对免疫系统和疫苗免疫效果具有调节作用。由于β-1，3/1，6-葡聚糖具有生物活性高、热力低等性能，可作为脂肪替代品、功能因子、膳食纤维等广泛应用于医药开发、保健食品、动物饲养等行业中，是一种值得研究和开发的功能性饲料添加剂。作者在介绍β-1，3/1，6-葡聚糖的来源、作用机理和生物活性的基础上，重点论述了其添加在畜禽日粮中对畜禽肠道的发育、生产性能、免疫功能的影响，同时对其应用时应注意的问题进行了阐述，旨在为β-1，3/1，6-葡聚糖在畜禽日粮中的应用奠定理论基础。     

Molecular approaches for enhancing sweetness in fruits and vegetables

The quality of fruits and vegetables is mainly dependant on the sweetness determined by the level of soluble sugars such as glucose, fructose and sucrose. Other fruit quality parameters include Brix content, acidity, aroma, color, size and shape. Total sugar content in fruits and vegetables is a function of genetic, nutritional, environmental and developmental factors. Understanding the factors controlling sweetness is important to design strategies for enhancing quality of fruits and vegetables. Modifying the activity of enzymes in carbohydrate metabolism such as sucrose synthase (SuSy), acid invertase, ADP-glucose pyrophosphorylase (AGPase), sucrose phosphate synthase (SPS) and sucrose transporters were found to influence carbohydrate partitioning and sucrose accumulation in sink tissues of several food crops. Plant based taste-modifying sweet proteins such as brazzein, cucurmin, mabinlin, monellin, miraculin, neoculin and thaumatin have potential application for developing transgenic plants to improve the sweetness and quality of fruits and vegetables. The present review envisages various cultural, breeding and molecular approaches used for enhancing sugar content and sweetness in fruits and vegetables.     

油菜短柄叶光合衰退及其对产量的影响

以中双9号、中油杂11号和华油杂14号油菜为材料,测定花后短柄叶光合速率及生理指标,探讨短柄叶光合衰退与叶绿素含量等因素之间的关系及其对油菜产量的影响。结果表明,油菜短柄叶光合速率随叶片衰老而逐渐降低,1,5-二磷酸羧化酶(RuBPCase)、可溶性蛋白和叶绿素含量,以及超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性均呈现出下降趋势,膜脂过氧化作用产物丙二醛(MDA)在叶片光合功能衰退过程中含量逐渐上升。油菜叶片光合衰退与RuBPCase含量、可溶性蛋白含量、叶绿素含量和SOD活性存在显著相关关系,表明光合速率的下降可能与油菜叶片衰老过程中N含量、叶绿素降解及蛋白质合成受抑制相关;SOD和CAT水平逐渐降低及MDA含量升高也表明,叶片衰老过程中氧自由基胁迫抑制光合作用。叶片光合作用与油菜单株产量的相关关系分析结果表明,油菜短柄叶光合速率,特别是短柄叶衰退的中后期光合速率是影响油菜产量的重要因素。     

腾冲红花油茶花蜜蛋白1,6-二磷酸果糖醛缩酶的鉴定、克隆与分析

为鉴定出花蜜中某些影响花蜜化学组成的功能蛋白,并探讨其在花蜜分泌及组分调控过程中的功能,以腾冲红花油茶为试验材料,利用MALDI-TOF/MS质谱分析方法初步鉴定出腾冲红花油茶花蜜中含有1,6-二磷酸果糖醛缩酶(FBP2),然后根据质谱得到的部分肽段序列设计特异性引物,采用SMART RACE方法克隆腾冲红花油茶花蜜FBP2全长cDNA,命名为CRFBPase (GenBank登录号:MG764086)。序列分析表明,该cDNA全长1 077 bp,编码358个氨基酸,分子质量为38 428.13 Da,等电点为7.56。经同源比对分析发现CRFBPase基因序列与普通油茶的FBP2基因序列同源性高达98.8%,同时酶活检测结果也进一步证实该花蜜中含有FBP2,且该酶的活性会随着花蜜分泌累积时间的延长而增加。实时荧光定量PCR结果显示,CRFBPase基因在开花后第5天的蜜腺和花部组织中均有一定量的表达且蜜腺中表达量最高,开花后第5天(泌蜜高峰期)蜜腺显著高于花蕾期。本研究结果为进一步揭示FBP2在植物花蜜形成、分泌及糖组分调控过程中的作用机制提供了一定的理论依据。     

猪肺炎支原体醛缩酶原核表达及致猪气管上皮细胞凋亡的研究

为了研究Ⅱ类果糖二磷酸醛缩酶(FBA)是否为猪肺炎支原体(Mycoplasma hyopneumoniae,Mhp)的毒力因子,并参与其致病作用,根据已发表的猪肺炎支原体醛缩酶全基因序列设计特异性的引物,以猪肺炎支原体168株基因组为模板,通过Overlap PCR点突变扩增醛缩酶基因。将FBA克隆至pET-28a(+)载体后进行序列测定和分析,结果表明,FBA基因全长864 bp。将构建的重组表达质粒pET-28a(+)/FBA转化至大肠埃希菌BL21(DE3),通过筛选获得阳性克隆,重组工程菌在IPTG诱导下成功获得表达融合蛋白猪肺炎支原体Ⅱ类果糖二磷酸醛缩酶(Mhp FBA),大小约为35 ku。纯化蛋白并接种于猪气管上皮细胞至FBA终浓度分别为0、10、50、100、150、200μg/mL,孵育24 h后进行上清中丙酮酸浓度的检测和细胞凋亡检测。结果上清中丙酮酸浓度随着FBA浓度的上升,出现先极显著上升(P<0.01)、后急剧下降的现象;细胞平均凋亡率依次分别为3.54%、10.91%、13.99%、27.19%、32.84%和40.71%,并随着FBA浓度的升高而升高,且相比阴性对照组差异均显著(P<0.05)或极显著(P<0.01)。成功构建了Mhp FBA重组菌,表达获得了Mhp重组蛋白FBA,该蛋白可能为猪肺炎支原体的毒力因子,参与猪肺炎支原体的致病作用,从而为进一步开展Mhp致病机理和药物研究奠定基础。     

中国明对虾FBA基因克隆及其在白斑综合征病毒感染中的表达及功能分析

醛缩酶(FBA)是糖酵解和糖异生中的关键酶,参与多种生物过程。本研究采用RACE技术,克隆获得中国明对虾(Fenneropenaeus chinensis)FBA基因(Fc FBA)的全长c DNA序列,并对其进行生物信息学分析。结果显示,中国明对虾Fc FBA基因的c DNA全长为2496 bp,其中,ORF长1098 bp,5~′UTR长79 bp,3~′UTR长1319 bp。完整的阅读框编码365个氨基酸,分子量为39.8 k Da,预测的理论等电点为6.6。同源性及系统进化分析表明,Fc FBA与节肢动物的FBA聚为一类,与卤虫(Artemia franciscana)、家蚕(Bombyx mori)、沙漠蝗(Schistocerca gregaria)的相似度分别是86%、79%和78%。荧光定量PCR结果显示,Fc FBA在肌肉中的相对表达量最高,肝胰腺中最低。WSSV感染后,该基因在鳃、肝胰腺和肌肉中呈现出不同的时空表达特点。ds RNA干扰24 h以后,抑制效率达到最大。与PBS对照组相比,Fc FBA干扰组(ds RNA组)加快了对虾染病后的死亡速度。本研究表明,Fc FBA基因可能参与了中国明对虾生物胁迫的应答反应。     

Insect-neuroactive substances in two species of the genus Liquidamber

Three insect-neuroactive substances from Liquidamber styraciflua and L formosana were isolated and characterized by spectral analyses as betulonic acid, 1-methoxy-9-caryolanol and eudesm-4(14)-ene-1,6-diol, respectively. ©1999 Society of Chemical Industry