玫瑰茄县浮细胞合成花青素的光效应研究

光照对悬浮培养的玫瑰茄细胞生物量无影响。随着光照强度增大,玫瑰茄细胞合成花青素的量增加,光照强度３１．０ｗ／ｍ＾２为饱和光照强度,超过该强度,玫瑰茄细胞合成花青素的量不再进一步增加;可见光中蓝光（４２０￣５３０ｎｍ）是促进玫瑰茄细胞合成花青素最有效单色光,光强为３０．０ｗ／ｍ＾２,接种量为０．２ｇ湿细胞的５０ｍｌ培养液经１６ｄ培养,花青素产量为８．９７ｍｇ／５０ｍｌ,高出相同光照强度全色光下的６．     

单色光玫瑰茄悬浮细胞合成花青素的影响

玫瑰茄悬浮细胞合成花青素受光调节。将不同的单色滤光膜覆盖在摇瓶表面,控制光照强度,判定了蓝光是促进玫瑰茄细胞产花青素的最有效单色光;红光和橙光无效;其它单色光随其波长接近蓝光,正交应增加。单色光对玫瑰茄细胞培养过程的其它参数,如ｐＨ、降糖、细胞生物量等影响不大。     

单色光对玫瑰茄悬浮细胞合成花青素的影响

玫瑰茄悬浮细胞合成花青素受光调节。将不同的单色滤光膜覆盖在摇瓶表面,控制光照强度,判定了蓝光是促进玫瑰茄细胞产花青素的最有效单色光;红光和橙光无效;其它单色光随其波长接近蓝光,正效应增强。单色光对玫瑰茄细胞培养过程的其它参数,如ｐＨ、降糖、细胞生物量等影响不大。     

主要营养成分对悬浮培养玫瑰茄细胞生长和花青素合成的影响

本文研究了培养基中碳源和氮源变化对悬浮培养玫瑰茄细胞生长和花青素合成的影响。在８种不同的碳源中,麦芽糖有利于花青素的积累,而蔗糖和葡萄糖适合细胞生长,并有较高的花青素产率。在１％～１０％蔗糖浓度范围内,４％浓度下细胞生长和花青素产率最高,而６％浓度下细胞花青素含量最高,高渗环境较有利于细胞花青素的积累。１３５ｍＭ的氮源总量已足够维持玫瑰茄细胞生长和花青素合成,氮源总量增加对细胞代谢有抑制作用。ＮＨ＋４对细胞有显著抑制作用。总量１３５ｍＭ,ＮＯ－３与ＮＨ＋４比例２５∶２和２３∶４时细胞生长和花青素合成最佳。     

悬浮培养玫瑰茄细胞的生长行为及动力学方程的建立

以培养多年的玫瑰茄（Hibiscus sabdariffaL.)白色细胞株为材料,在以3％蔗糖为碳源,植物生长调节剂为4．5μmol/L2,4-D（2,4－二氯苯氧乙酸）,2．3μmol/L Kt（Kinetin,细胞激动素）的悬浮培养条件下,研究了玫瑰茄细胞的生长及泛醌积累动态、培养液PH值的变化以及培养过程中玫瑰茄细胞对碳源、氮源、磷源的消耗动态,并以此数据建立了悬浮培养玫瑰茄细胞的生长、蔗糖消耗、泛醌形成的非结构动力学模型,经验证表明,模型与实际情况有较高的拟合度。     

不同碳源对悬浮培养玫瑰茄细胞主要基质消耗的影响

采用了蔗糖、葡萄糖、可溶性淀粉3种不同碳源,考察了在摇瓶培养过程中,这3种碳源对维持悬浮培养过程中玫瑰茄细胞的生长及主要基质消耗的影响。结果表明,蔗糖和葡萄糖在这些方面的表现基本相同,而可溶性淀粉由于植物细胞对其没有有效的水解利用手段,因而不能支持植物细胞的生长,从基质消耗水平上来看,也明显慢于上述两类碳源。     

玫瑰茄中营养元素的分析研究

通过原子吸收火焰光度法对玫瑰茄中的营养元素进行测定分析,结果表明：玫瑰茄中富含钙、镁营养元素,富含铁、锰、锌微量元素,从而为玫瑰茄的食用、医用价值及保健功能提供科学理论依据。     

中间产物对玫瑰茄培养细胞合成花青苷的影响

用Ｂ５培养基悬浮培养产色素的玫瑰茄培养细胞,培养１３天时,花青苷产量最高,为０．２５ｇ／Ｌ。培养基中添加终浓度为１０＾－６ｍｏｌ／Ｌ的外源Ｌ－Ｐｈｅ能够显著地增加产色素细胞花青苷的积累量。浓度为１０＾－７ｍｏｌ／Ｌ的槲皮素,可使悬浮培养的玫瑰茄细胞花青苷产量提高１．３倍,无论是Ｌ－Ｐｈｅ还是槲皮素均不能启动不产色素的细胞系产花青苷。     

不同氮湖对悬浮培养玫瑰茄细胞的生长及硝态氮同化指征的影响

本文通过改变培养基中的氮源组成,来研究氮源的变化以悬浮培养玫瑰茄细胞的生长及硝态所同化指征的影响。实验表明,仅在含ＮＯ＾－３的培养基中能检测到硝酸还的酶活力,而仅含ＮＨ＾＋４的配方不能检测到该酶活力,表明硝酸还原酶是底物诱导酶。     

玫瑰茄培养细胞中花色苷积累的代谢调节

调整培养基组成是提高玫瑰茄（ＨｉｂｉｓｃｕｓｓａｂｄａｒｉｆｆａＬ．）培养细胞花色苷产量的有效途径之一。改变Ｂ５培养其中钙、锰、铜等离子的浓度,增加了培养细胞花色苷的积累量,其中又以锰离子最为有效。其浓度为６＊１０＾４ｍｏｌ／Ｌ时,花色苷磁量达０．３４１ｇ／Ｌ,是对照的２．８２倍。     

高产花色苷玫瑰茄细胞系的筛选

花色苷在植物中呈现粉红、红、紫红、紫等颜色,可以用作食品、药品及化妆品的着色剂,亦有药用价值。作为食品添加剂,颜色较合成色素自然,且安全无毒性。早在１９８７年,Ｍｉｚukami^[1]就建议用植物细胞培养物生产花色苷类代替合成色素。所有的植物培养细胞都是异源性的。各细胞之间产花色苷的能力相差很大^[2].因为产花色苷的细胞系带有颜色标记,所以容易识别并通过肉眼选择即可获得高产花色苷的细胞系。筛选的方法很多,如平板饲喂法^[3]、小细胞团法^[4]、细胞块法^[5]、肉眼观察直接挑选法及细胞分栋器法^[6]等。高产系花色苷的含量可增加几倍到几十倍,而且产量稳定。本文采用平板法及小细胞团法筛选高产花色苷的玫瑰茄(Hibiscus sabdariffa L.)细胞系。     

Effect of light irradiation on anthocyanin production by suspended culture of Perilla frutescens

After a series of experiments on photoperiodicity and light intensity under daylight supplied by an ordinary fluorescent lamp in cultivations using a flask and a roux bottle, it was found that irradiation at 27.2 W/m(2) for the whole period was effective for anthocyanin production by a suspended culture of Perilla frutescens (shiso). A high amount of anthocyanin pigments, 3.0 g/L, was obtained in a bubble column bioreactor after 10 days of cultivation at an aeration rate of 0.1 vvm with light irradiation at 27.2 W/m(2), while 2 g/L was obtained at 13.6 W/m(2) and very little at 54.4 W/m(2). A high amount of anthocyanin pigments, 2.9 g/L, was also produced using an aerated and agitated bioreactor at an agitation speed of 130 rpm, an aeration rate of 0.1 vvm and light irradiation intensity of 27.2 W/m(2). The amount of anthocyanin produced was more than twice that without light irradiation, Keeping the other cultivation conditions the same. The results obtained also showed that the amount of anthocyanin pigment accumulated in a shake flask could be rather well reproduced in bioreactors for both aerated culture, and aerated and agitated culture, by improving the conditions of light irradiation, which conspicuously affects metabolite formation.     

Anthocyanin production in callus cultures of roselle (Hibiscus sabdariffa L.)

Callus tissues derived from seedlings of roselle (Hibiscus sabdariffa L.) were shown to produce two cyanidin glycosides as major anthocyanin pigments. Both callus growth and anthocyanin synthesis were remarkably stimulated by 2,4-dichlorophenoxyacetic acid. The highest anthocyanin yield was observed when 1 M 2,4-D in combination with 0.1–1 M kinetin was supplemented to the culture medium. In contrast, gibberellic acid showed inhibitory effect on anthocyanin production.Abbreviations LS Linsmaier and Skoog - 2,4-D 2,4-dichlorophenoxyacetic acid - NAA 1-naphthaleneacetic acid - IAA indole-3-acetic acid - IBA indole-3-butyric acid - GA gibberellic acid     

Antihypertensive effect of roselle (Hibiscus sabdariffa) calyx infusion in spontaneously hypertensive rats and a comparison of its toxicity with that in Wistar rats

The LD₅₀ of roselle calyx extract and its effect on blood pressure were determined. The LD₅₀ was found to be above 5000 mg kg⁻¹. Roselle calyx infusion was found to lower significantly (p<0·05) both systolic and diastolic pressure in spontaneously hypertensive and normotensive Wistar–Kyoto rats at tested doses of 500 and 1000 mg kg⁻¹ body weight. The reduction in blood pressure in both groups was positively correlated with weight. Continuous consumption of the infusion at 1000 mg kg⁻¹ was discovered to lead to sudden death in spontaneously hypertensive rats but not in Wistar‐Kyoto rats. Water intake was not significantly different (p>0·05) in the control groups of the two strains of rats used, neither was there a significant difference in their urine output. The water intake in the treated spontaneously hypertensive and normotensive rats was not different from the corresponding control groups. However the urine output of the treated spontaneously hypertensive rats was significantly higher. A significant decrease in serum creatinine, cholesterol, and glucose in the treated rats compared with the control as well as a significant increase in serum uric acid was observed. The serum proteins (albumin and total protein) in the treated rats when compared with the control groups was not changed significantly. Copyright © 1999 John Wiley & Sons, Ltd.     

Prevailing insects in Roselle plants,Hibiscus sabdariffa L., and their efficiency on pollination

Field surveys of Roselle plant fields were conducted during two successive seasons of 2006 and 2007. The total counted insects on Roselle, Hibiscus sabdariffa attained 16 species belong to seven insect Orders [(Hemiptera (3 species), Homoptera (1 species), Lepidoptera (3 species), Neuroptera (1 species), Coleoptera (3 species), Diptera (1 species), Hymenoptera (4 species)]. Variable numbers of insects were counted on different Roselle varieties and in the two seasons. The most prevailing insect species were the piercing sucking pests [Empoasca spp.; Spilostethus longulus Dallas; Nezara viridula L. and Oxycarenus hyalinipennis (Costa)] and the predators [Polistes sp., Coccinella undecimpunctata and Scymnus syriacus]. Honeybee (Apis mellifera L.) proved to be the most abundant pollinator, averaging 13, 17, 18 individuals/25double strikes in cultivations of Sudani, Masri and White varieties of Roselle plants during flowering of the first season respectively. Similar results were also obtained in the second season. The highest number of honeybees was recorded around 10 am to midday during the flowering stage for the two seasons. It was obvious that the effects of the two hydrothermal, factors temperature and relative humidity, had slight or negligible effects on the occurrence of insect pollination throughout the two seasons of the study, while statistical analysis of the obtained results showed that the productivity of Roselle plant (mean number of seeds and mean weights of seeds) were significantly affected by pollinators.