二苯胺棉酚中棉酚的含量测定

用紫外分光光度法测定二苯胺棉酚中棉酚的含量。方法回收率平均达 93.6 %以上 ,变异系数 ( CV )为1.2 %。该方法准确 ,重现性好 ,可作为二苯胺棉酚质量监控方法。     

棉籽脱酚工艺的研究

本文研究了分步浸油和萃取棉酚的工业化生产工艺，解决了生产过程中遇到的设备和工艺问题。采用该工艺生产的脱酚棉籽蛋白中游离棉酚含量小于0．045％、残油小于1．5％、蛋白含量大于50％。     

脱除游离棉酚乳酸菌的分离鉴定及益生性能研究

从藜麦茎叶样品中筛选分离出乳酸菌株,测定菌株脱除游离棉酚能力,通过人工胃液、人工肠液耐受实验、抑菌实验、抗生素敏感实验及表面特性研究菌株各项性能,并通过生理生化及16S rDNA法对菌株进行鉴定。结果表明,筛选出1株高效游离棉酚脱除乳酸菌LR002,其游离棉酚脱除率为62.87%,该菌株具有高效的耐人工胃液、肠液效果,且具有广谱抑菌作用,对抗生素不敏感,经鉴定该菌株为植物乳杆菌（Lactobacillus plantarum）。本研究旨在筛选出高效游离棉酚脱除乳酸菌菌株,为棉粕在畜禽生产中的合理利用提供参考。     

低酚棉中苯丙氨酸解氨酶活力与某些含氮化合物的变化（简报）

低酚棉植株中苯丙氨酸解氨酶(PAL)活力比有酚棉中的低。在营养生长期,低酚棉叶片含氮量基本保持恒定,有酚棉则呈下降趋势。低酚棉棉籽中的粗蛋白含量随生育期的延续呈上升趋势,而有酚棉中的则基本保持恒定。     

高效降解棉酚菌株的分离鉴定及诱变选育

作为一种蛋白资源, 棉籽粕因其含有毒素——游离棉酚限制了其在饲料工业中的应用。为获得能高效降解棉籽粕中棉酚的菌株, 以醋酸棉酚为唯一碳源培养基从16份样品中筛选出一株高效降解棉酚菌株(Y-2), 经生理生化及18S rDNA鉴定为Pichia guilliermondii, 此菌种为非致病酵母, 且首次报道用于棉酚的降解, 在工业生产中具有潜在的应用前景。通过紫外诱变获得一株棉酚降解率更高的突变株YUV-51。通过对突变株YUV-51发酵温度、时间及接种量的初步优化, 获得其固态发酵优化条件: 30 °C培养48 h, 接种量0.025 g湿菌体/g棉籽粕, 初始水分含量50%。为避免游离棉酚在前处理中大量降解棉籽粕, 不进行湿热处理, 经接种发酵后脱毒率可达到58%。这使微生物脱毒在实际生产中应用成为可能。     

高效棉酚降解枯草芽孢杆菌M-4菌株的常压室温等离子体诱变及发酵工艺优化

【背景】棉粕中游离棉酚的存在制约了棉粕作为饲料蛋白源的利用,棉酚的微生物降解问题成为研究热点。本实验室前期发现枯草芽孢杆菌(Bacillus subtilis) M-4菌株具有较强的降解棉酚能力,而且已经应用于棉粕脱毒工业。【目的】进一步提高枯草芽孢杆菌M-4菌株降解棉粕中棉酚的能力,扩大棉粕在养殖业的应用领域。【方法】采用常压室温等离子体诱变(atmospheric and room temperature plasma,ARTP)技术对菌株M-4进行诱变,以液体培养条件下的棉酚降解率为初筛指标,筛选获得正向突变株。以棉粕固体发酵条件下的棉酚降解率为复筛指标,测定初筛获得的正向突变株实际降解棉粕中游离棉酚的能力。以棉酚残存量和棉酚降解率为检测指标,采用单因素试验优化固体条件下棉粕发酵条件,获得突变株发酵棉粕的最适工艺参数。【结果】初筛获得正向突变株19株。复筛得到一株高效的突变株MY-4-17,在棉粕固体发酵条件下其棉酚降解率高达97.15%,比菌株M-4的棉酚降解率提高了2.55%。经过5次传代培养,突变株MY-4-17的遗传稳定性良好。突变株MY-4-17发酵棉粕的最适工艺参数为...     

棉叶中棉酚的快速提取与测定方法研究

棉酚(G)及其相关物甲氧基半棉酚(DHG)、半棉酚酮(HGQ)、半棉酚(HG)、杀实夜蛾素(H1-4)等是棉花中重要的抗虫性萜烯类次生物质.利用高效液相色谱法(HPLC)对棉酚及其相关物进行了分离,测定了棉叶中的棉酚含量,讨论了不同的提取方法和测定条件对结果的影响,给出了一套简便、快速的分析测试方法,同时与紫外-可见分光光度法(苯胺法)的结果进行了对比,认为对于棉花的抗虫性的研究来说,HPLC是比较适宜的方法.     

棉族色素腺体的利用

棉族(Tribe Gossypieae) 是锦葵科(Malvaceae)内一个分类单位。色素腺体是棉族植物所特有的一个重要特征,迄今为止,在其他植物中尚未发现色素腺体的存在。色素腺体具有一定的形态,含有多种有毒的有色化学物质,其中棉酚是主要的化学成分,占色素腺体重量的39—50%,它是棉族这一类群最显著最一致的特征。棉酚对人和非反刍动物有毒,这限制了对棉籽这一富含蛋白质和脂     

旋光异构体棉酚对精子脱氢酶的影响

本研究采用组织化学方法,比较了左旋(-)、右旋(+)及消旋(±)棉酚,对人和豚鼠离体精子以及喂服三型棉酚4或8周的大鼠精子五种脱氢酶(乳酸脱氢酶LDH、苹果酸脱氢酶MDH、异柠檬酸脱氢酶ICDH,甘油3-磷酸脱氢酶G3-PDH和α-羟戊酸脱氢酶VDH)的影响。并将酶活性用细胞光度计做了定量测定。体外实验结果表明:相同剂量的(一)棉酚与(+)棉酚相比,前者对酶活性、精子形态及线粒体结构影响更加明显。两种棉酚之间的酶活性改变有统计学意义。在体实验中,比较三型棉酚对动物健康状况、精子数量、活动率和形态变化的影响,发现(+)棉酚不是一种有效的抗生育制剂。 (±)和(一)棉酚在产生抑精效应的同时,(±)棉酚毒性较大,表现为动物体重下降,精子数量甚少。(一)棉酚具有两倍于(±)棉酚的抑精作用,是一种高效低毒的酚类化合物。本文还就线粒体结构损伤与酶活性改变的关系,以及棉酚的作用机理作了讨论。     

棉酚对Jurkat T细胞增殖与凋亡的影响

研究棉酚(gossypol)对Jurkat T细胞增殖和凋亡的影响及可能机制.将不同浓度的棉酚作用于Jurkat T细胞,用MTS比色法检测细胞存活率;以膜联蛋白V-PE染色分析细胞凋亡;用Hoechst33342染色观察核形态:用荧光染料Mitocapture结合流式细胞术和激光共焦显微镜检测线粒体跨膜电位变化.结果显示棉酚作用Jurkat T细胞24 h、48 h、72 h,其IC50值分别为77.2μmol/L、57.3 μmol/L、23.3 μmol/L,对细胞的抑制作用与药物存在时间-剂量依赖关系;终浓度为8、16、32 μmol/L的棉酚处理24 h后的细胞凋亡率分别为5.30%、15.20%、51.19%,对照组凋亡率为3.43%;高浓度棉酚作用后大量细胞核呈现染色质固缩、核碎裂和致密浓染等凋亡特征;随着浓度增加细胞线粒体跨膜电位明显降低.研究表明棉酚能有效抑制Jurkat T细胞增殖和诱导其发生凋亡,并呈现出时间-剂量依赖关系,其诱导凋亡的作用可能依赖于线粒体途径.     

RNAi‐mediated Ultra‐low gossypol cottonseed trait: performance of transgenic lines under field conditions

Cottonseed remains a low‐value by‐product of lint production mainly due to the presence of toxic gossypol that makes it unfit for monogastrics. Ultra‐low gossypol cottonseed (ULGCS) lines were developed using RNAi knockdown of δ‐cadinene synthase gene(s) in Gossypium hirsutum. The purpose of the current study was to assess the stability and specificity of the ULGCS trait and evaluate the agronomic performance of the transgenic lines. Trials conducted over a period of 3 years show that the ULGCS trait was stable under field conditions and the foliage/floral organs of transgenic lines contained wild‐type levels of gossypol and related terpenoids. Although it was a relatively small‐scale study, we did not observe any negative effects on either the yield or quality of the fibre and seed in the transgenic lines compared with the nontransgenic parental plants. Compositional analysis was performed on the seeds obtained from plants grown in the field during 2009. As expected, the major difference between the ULGCS and wild‐type cottonseeds was in terms of their gossypol levels. With the exception of oil content, the composition of ULGCS was similar to that of nontransgenic cottonseeds. Interestingly, the ULGCS had significantly higher (4%–8%) oil content compared with the seeds from the nontransgenic parent. Field trial results confirmed the stability and specificity of the ULGCS trait suggesting that this RNAi‐based product has the potential to be commercially viable. Thus, it may be possible to enhance and expand the nutritional utility of the annual cottonseed output to fulfil the ever‐increasing needs of humanity.     

In situ alkaline transesterification of cottonseed oil for production of biodiesel and nontoxic cottonseed meal

The production of fatty acid methyl ester (FAME) by direct in situ alkaline-catalyzed transesterification of the triglycerides (TG) in cottonseeds was examined. The experimental results showed that the amount of cottonseed oil dissolved in methanol was approximately 99% of the total oil and the conversion of this oil could achieve 98% under the following conditions: less than 2% moisture content in cottonseed flours, 0.3-0.335mm particle size, 0.1mol/L NaOH concentration in methanol, 135:1 methanol/oil mole ratio, 40 degrees C reaction temperature and 3h reaction time. Further, the effects of co-solvent petroleum ether and methanol recycling on the cottonseed oil extraction and conversion were also investigated. The use of alkaline methanol as extraction and reaction solvent, which would be useful for extraction oil and gossypol, would reduce the gossypol content in the cottonseed meal. The free and total gossypol contents in the cottonseed meal obtained from in situ alkaline transesterification were far below the FAO standard. And the nontoxic cottonseed meal could be used as animal protein feed resources.     

In vitro inhibitory effect of gossypol from gossypol-acetic acid, and (+)- and (-)-isomers of gossypol on the growth of Edwardsiella ictaluri

AIM: This study was conducted to evaluate the toxic effect of gossypol from gossypol-acetic acid, and (+)- and (-)-isomers of gossypol on the growth of Edwardsiella ictaluri. METHODS AND RESULTS: Inhibitory effect of various concentrations of gossypol on the growth of E. ictaluri was determined. Bacterial recovery was performed by preincubation of bacteria in medium containing various concentrations of gossypol and subsequent activation of bacteria by inoculating on gossypol-free plates. Concentrations of racemic gossypol, (+)-gossypol and (-)-gossypol of 1.5 microg ml(-1) or higher significantly reduced the number of bacterial colonies compared with that of the control. The growth of E. ictaluri was completely inhibited on agar plates supplemented with 3 microg ml(-1), regardless of the forms of gossypol. The inhibitory effect of (+)-gossypol was higher than that of (-)-gossypol or gossypol-acetic acid. Recovery of E. ictaluri was <50% for all three forms of gossypol at concentrations of 5 microg ml(-1). Bacterial recovery remained relatively constant (6.5%) at gossypol concentrations from 10 to 100 microg ml(-1). Complete killing of E. ictaluri was not reached at gossypol levels up to 100 microg ml(-1). CONCLUSION: Gossypol-acetic acid, and (+)- and (-)-optical isomers have anti-bacterial effect against E. ictaluri. The results suggest the action is bacteriostatic rather than bactericidal. SIGNIFICANCE AND IMPACT OF THE STUDY: The therapeutic effect of gossypol against E. ictaluri may be useful in controlling enteric septicaemia of catfish.     

Temporal allocation of metabolic tolerance in the body of beet armyworm in response to three gossypol-cotton cultivars

The nutrient composition and enzyme activities in larvae of the beet armyworm, Spodoptera exigua (Hübner), fed on high, medium or low gossypol cotton cultivars were examined at different time intervals. Significantly lower free fatty acid was observed in larvae fed for 6 h on high gossypol ‘M9101’ compared to larvae fed on the low (ZMS13) and intermediate (HZ401) gossypol cultivars. Significantly higher trypsin activity was observed in larvae fed on high gossypol ‘M9101’ for 24 h compared to those fed for 1, 4 and 6 h. Significantly higher catalase and total superoxide dismutase enzyme activities were observed in larvae of S. exigua fed on high gossypol ‘M9101’ compared with low gossypol cultivars ‘ZMS13’ and ‘HZ401’ for 1, 4, 6 and 24 h. However, significantly lower carboxylesterase and acetylcholinesterase enzyme activities were found in larvae fed on high gossypol ‘M9101’ compared with the other cultivars for 1, 4, 6 and 24 h. The interaction between cotton variety and beet armyworm infestation time significantly affected the carboxylesterase enzyme activity in S. exigua. The characterization of the effects of plant allelochemicals on herbivorous larvae is important for aiding understanding of plant-insect interaction as well as in devising solutions to pest problems by breeding plant resistance, identifying metabolic targets for insecticide development, etc.     

Effect of gossypol on intracellular Ca2+ regulation in human hepatoma cells

Gossypol is a natural toxicant present in cottonseeds, and is hepatotoxic to animals and human. The effect of gossypol on cytosolic free Ca2+ levels ([Ca2+]i) in HA22/VGH human hepatocytes was explored using fura-2 as a fluorescent Ca2+ indicator. Gossypol increased [Ca2+]i in a concentration-dependent manner with an EC50 value of 2 microM. The Ca2+ signal was reduced by removing extracellular Ca2+ or by 10 microM La3+, but was not affected by nifedipine, verapamil or diltiazem. Pretreatment with 1 microM thapsigargin (an endoplasmic reticulum Ca2+ pump inhibitor) to deplete the endoplasmic reticulum Ca2+ partly reduced 10 microM gossypol-induced Ca2+ release; and conversely pretreatment with gossypol abolished thapsigargin-induced Ca2+ release. The Ca2+ release induced by 10 microM gossypol was not changed by inhibiting phospholipase C with 2 microM U73122 or by depleting ryanodine-sensitive Ca2+ stores with 50 microM ryanodine. Together, the results suggest that in human hepatocytes, gossypol induced a [Ca2+]i increase by causing store Ca2+ release from the endoplasmic reticulum in a phospholipase C-independent manner, and by inducing Ca2+ influx.     

EFFECTS OF GOSSYPOL ON GROWTH OF THE COTTON BOLLWORM AND DEVELOPMENT OF ITS PARASITOID CAMPOLETIS CHLORIDEAE*

Abstract The effects of gossypol, an important allelochemical in cotton, on growth of the cotton bollworm Helicoverpa armigera (Hubner) and development of its endoparasitoid Campoletis chlorideae (Uchida) in Huanghe River Valley were studied. Growth of H. armigera larvae was accelerated by adding 0.1% gossypol in the artificial diet, causing 10.75% reduction of the vulnerable period to C. chlorideae while the suppression activity of 0. 5 % gossypol to H. armigera larvae prolonged the vulnerable period by 28.15%. Negative effects of gossypol on the development of the parasitoid were demonstrated by using the artificial diet and cotton varieties WD-151 (glandless) and HG-BR-8 (glanded). Gossypol at 0.1% concentration in artificial diet did not remarkably increase the body weight of adult wasp, but significantly extended the egg-larval time and shortened the pupal time; 0. 5% gossypol and HG-BR-8 remarkably reduced the body weight of adult wasp, also significantly prolonged the egg-larval period and decreased the pupal period. Finally, the coordinate application of high gossypol resistant cotton varieties and C. chlorideae in H. armigera control is discussed.