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．     

青蒿毛状根合成青蒿素的培养条件研究

对影响青蒿（ＡｒｔｅｍｉｓｉａａｎｎｕａＬ．）毛状根生长及青蒿素合成的培养条件进行了研究,确定最适的培养条件为：初始ｐＨ５．８～６．０,摇瓶转速１３０～１５０ｒ／ｍｉｎ,摇瓶装液量体积分数为２５％,光照周期为１６ｈ／ｄ,温度为３０℃。在此条件下,经过２５ｄ培养获得青蒿素产量为２２３．３ｍｇ／Ｌ。     

黄花蒿培养细胞中青蒿素合成代谢的体外调节

黄花蒿培养细胞通过两步培养积累青蒿素．第１步在含有０．２～０．４ｍｇ／Ｌ６－苄基氨基嘌呤（６－ＢＡ）和３～４ｍｇ／Ｌ吲哚乙酸（ＩＡＡ）的Ｎ６培养基中进行细胞的增殖培养,第２步将培养好的细胞转入含０．２～０．４ｍｇ／Ｌ６－ＢＡ和０．２～０．４ｍｇ／ＬＩＡＡ的改良Ｎ６培养基中进行青蒿素的合成．青蒿素的合成量为１９０μｇ／ｇ干细胞左右．当在第２步培养中加入青蒿素合成前体青蒿酸,青蒿素合成量比仅靠激素诱导提高了３倍多．青蒿素的合成途径是植物固醇合成途径的分支途径,当在青蒿素合成过程即第２步培养中加入固醇生物合成抑制剂双氯苯咪唑和氯化氯胆碱处理,可使代谢向合成青蒿素的方向移动,青蒿素合成量明显提高．经２００ｍｇ／Ｌ氯化氯胆碱处理２ｄ,黄花蒿细胞合成青蒿素量为３７２μｇ／ｇ干细胞;经２０ｍｇ／Ｌ双氯苯咪唑处理４ｄ,黄花蒿细胞合成青蒿素量为１５４０μｇ／ｇ干细胞,比靠激素诱导提高了８倍多,与诱导脱分化细胞的黄花蒿叶中所含的青蒿素（３０００μｇ／ｇ干细胞）处于同一个数量级．以上结果表明：在通过植物激素调节可以合成青蒿素的黄花蒿培养细胞中,缺乏青蒿素合成前体是青蒿素合成量低的重要原因．因此,在青蒿素合成的过程中通过体外调节,     

中麻黄悬浮培养体系的建立

本文用中麻黄无菌苗为外植体,其切段培养在附加２ｍｇ／Ｌ２,４－Ｄ和０．５ｍｇ／Ｌ　６　ＢＡ的ＭＳ培养基上,全部脱分化形成白色疏松愈伤组织。愈伤组织继代培养于ＭＳ＋０．５ｍｇ／Ｌ２,４－Ｄ＋０．２ｍｇ／Ｌ６ＢＡ＋０．２ｍｇ／Ｌ　ＮＡＡ＋４％蔗糖的培养某上。以继代培养愈伤组织为材料进行悬浮培养,培养基为附加０．２ｍｇ／Ｌ２,４－Ｄ＋０．１ｍｇ／Ｌ６ＢＡ＋０．１ｍｇ／ＬＮＡＡ＋２％蔗糖的ＭＳ液体培养基,得到分散性好,细胞形状接近圆形,细胞大小均一,细胞团多由２－３０个细胞组成的悬浮培养体系。第三代悬浮培养细胞增长率为０．３５ｇ·ｆｗ／２０ｍｌ·ｄ,细胞有丝分裂指数为１１．２％。条件培养和高密度接种可缩短延迟期,条件培养不能提高分裂指数,１ｇ／１０ｍｌ接种密度可使分裂指数提高至２１．２％。     

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。     

丹心Ⅲ号和丹心V号对血小板聚集功能的影响

本文观察了丹心Ⅲ号和丹心Ｖ号对血小板聚集功能的影响,实验结果：丹心Ⅲ号和丹心Ｖ号在体外均可显著抑制ＡＤＰ和花生四烯酸诱导的人血小板聚集,其ＩＣ５０分别为１．６０５ｍｇ／ｍｌ,２．５８９ｍｇ／ｍｌ,８．４ｌ６ｍｇ／ｍｌ和６．６０６ｎｇ／ｍｌ;在体内可抑制连续给药（３５０一７００ｍｇ／ｌｋｇ．ｄ）１０ｄ大鼠血小板对ＡＤＰ和花生烯酸诱导的血小板聚集,但对凝血酶诱导的血小板聚集无明显抑制作用。上述结果提示丹心Ⅲ号和丹心Ⅴ号对血小板聚集功能具有明显的抑制作用。     

影响大麦叶肉原生质体分离和培养的一些因子（简报）

从大麦幼苗分离叶肉原生质体，５ｄ苗龄的较４ｄ及６ｄ以上苗龄的得率高。提高Ｃａ＾２＋浓度有利于原生质体的分离，浓度为１０ｍｍｏｌ／Ｌ时的得率最高。在以添加０．５ｍｇ／Ｌ２，４－Ｄ、１．０ｍｇ／Ｌ ＮＡＡ及０．５ｍｇ／Ｌ ＺＴ的改良ＭＳ培养基和微弱的光照条件下，原生质体能持续分裂，并形成小细胞团。     

玉米雌穗离体培养诱导孤雌生殖结实

离体条件下,用１６种不同的玉米材料未授粉的雌穗进行整个直插或切段直插培养。培养在２８℃培养箱和室内散射光下,其培养基为：ＭＳ＋０．１ｍｇ／Ｌ２,４－Ｄ＋２ｍｇ／ＬＮＡＡ＋２ｍｇ／ＬＫＴ＋５００ｍｇ／ＬＬＨ＋４％蔗糖＋０．６％琼脂（ＰＨ５．８）和ＭＳ＋０．０５ｍｇ／Ｌ２,４－Ｄ＋２ｍｇ／ＬＮＡＡ＋２ｍｇ／ＬＫＴ＋４％蔗糖＋０．６％琼脂（ＰＨ５．８）。从未授粉的雌穗（成熟胚囊期）获得结实。这些种子萌发     

菜心下胚轴原生质体培养和植株再生

以萌发３—４ 天（长约４ ｃｍ ）的菜心（Ｂｒａｓｓｉｃａ ｃａｍｐｅｓｔｒｉｓ ｖａｒ．ｐａｒａｃｈｉｎｅｓｉｓ）无菌苗苍白下胚轴为材料,酶解分离原生质体。经纯化的原生质体,在含０．５ ｍ ｇ／ＬＺＴ、０．５ ｍ ｇ／Ｌ２,４－Ｄ、１．０ ｍ ｇ／ＬＮＡＡ 和０．４ ｍ ｏｌ／Ｌ葡萄糖的Ｋ８ｐ 培养基中,进行微滴培养。在起始培养１４—１８小时,原生质体再生新的细胞壁。３６ 小时再生细胞开始第一次分裂。第三天分裂细胞频率可达３５％ 。培养第８—９ 天,可见含８—１６个细胞的小细胞团,植板率为１５％ —１８％ 。３ 周后将发育成直径为２ ｍ ｍ 的白色小愈伤组织,转到含０．３ ｍ ｇ／Ｌ ２,４－Ｄ并用ｇｅｌｒｉｔｅ半固化的培养基上,增殖成４—５ ｍ ｍ 直径的愈伤组织。在ＭＳ＋ ３．２（或１．６） ｍ ｇ／Ｌ ＢＡ＋ １．６（或０．８） ｍ ｇ／ＬＺＴ＋ ０．０１ ｍ ｇ／Ｌ ＮＡＡ＋ ０．１ ｍ ｇ／ＬＧＡ３ 和０．２％ 蔗糖的分化培养基上,获得芽的分化。切下约２ ｃｍ 长的芽苗,转移到含０．２ ｍ ｇ／ＬＩＡＡ 和２％ 蔗糖的培养基上,生根形成完整植株     

杨树新品种叶肉原生质体培养和植株再生

从１ 个月龄的ＮＬ－８０１０６ 杨（Ｐｏｐｕｌｕｓｄｅｌｔｏｉｄｅｓ×Ｐ． ｓｉｍ ｏｎｉｉ）无菌苗叶片分离得到大量原生质体,纯化后其原生质体产量为４×１０７／ｇ ｆｒ．ｗ ｔ． 纯化的原生质体在含２,４－Ｄ ２ ｍ ｇ／Ｌ、ＮＡＡ ０．５ ｍ ｇ／Ｌ和ＫＴ ０．５ ｍ ｇ／Ｌ的ＫＭ８ｐ 和ＭＳ培养基中进行高密度液体浅层培养,渗透势为０．４０ ｍ ｏｌ／Ｌ的ＫＭ８ｐ 培养基中原生质体分裂频率最高． 培养第５ 天观察到第一次细胞分裂,培养１０ ｄ 的分裂频率为４．５％ ,１２ 周内可形成大量的细胞团和小愈伤组织． ＮＬ－８０１０６杨叶肉原生质体在富含有机氮并以葡萄糖为碳源的培养基中具有较高的分裂频率和植板率．小愈伤组织在ｇｅｌｒｉｔｅ 固化的ＮＬＺ１ 培养基上增殖生长,３ 周后形成４—６ ｍ ｍ 结构紧密的鲜红色愈伤组织,转至ＮＬＦ分化培养基,分化成苗率为１００％ ． 待芽伸长到３ ｃｍ 时,从基部切下转至１／２ ＭＳ培养基上诱导生根,形成完整植株     

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。     

Anthocyanin accumulation and changes in activities of phenylalanine ammonia-lyase and chalcone synthase in roselle (Hibiscus sabdariffa L.) callus cultures

Time-course changes in anthocyanin accumulation, phenylalanine ammonia-lyase activity and chalcone synthase activity were examined in roselle callus tissues incubated under different culture conditions. Phenylalanine ammonia-lyase activity was not affected by either the kind of auxin supplemented to the medium or light regime. In contrast, chalcone synthase activity was markedly suppressed when the callus was cultured with a medium containing indole-3-acetic acid instead of 2,4-dichlorophenoxyacetic acid (2,4-D) or in the dark. The results imply that in roselle callus cultures chalcone synthase plays a more important role in anthocyanin biosynthesis regulated by 2,4-D and light irradiation than phenylalanine ammonialyase.Abbreviations LS Linsmaier and Skoog - 2,4-D 2,4-dichlorophenoxyacetic acid - IAA indole-3-acetic acid - PAL phenylalanine ammonia-lyase - CHS chalcone synthase     

Some Factors Affecting the Anthocyanin Formation by Populus Cells in Suspension Culture

In order to elucidate the mechanism of anthocyanin formation by Populus cells in suspension culture, the favourable conditions for anthocyanin formation were investigated. The influence of some factors affecting the anthocyanin formation, i.e. light, sucrose and riboflavin etc. were also examined. Light irradiation and high sucrose concentrations brought about a marked increase of PAL activity, which increased rapidly at the lag phase preceding the anthocyanin formation. The effect of blue light on anthocyanin formation was markedly superior to other kinds of monochromatic light (green and red) or white light. Riboflavin was effective only under light exposure. It was inferred that light, sucrose, riboflavin and PAL activity etc. were closely related with the anthocyanin formation. Especially, light and sucrose cooperated in the increase of PAL activity which was a key enzyme in the biosynthesis of anthocyanin.     

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。     

Relative effectiveness and interaction of ultraviolet-B, red and blue light in anthocyanin synthesis of apple fruit

The effect of light on anthocyanin production in apple ( Malus pumila Mill. cv. Jonathan) skin disks was investigated, with prolonged irradiation from different light sources. High fluence rates of white light provided from a xenon lamp were unable to produce large amounts of anthocyanin, and anthocyanin production became saturated at about 30 W m⁻². When UV-B light, provided by a fluorescent lamp which had an emission peak at 312 nm, was combined with the white light, anthocyanin production was synergistically stimulated and increased up to the highest fluence rates of white light tested (44 W m⁻²). This UV-B light was more effective than red and blue light provided from fluorescent lamps, but anthocyanin production became saturated at about 1.7 W m⁻². However, simultaneous irradiation with red and UV-B light had a synergistic effect. UV-B light was also effective in increasing anthocyanin production in whole fruit. Therefore this synergism seemed to have an important role in the development of the desirable red skin color under field light conditions. The results of aminoethoxyvinylglycine treatment suggested that ethylene was not involved in the stimulative effect of UV-B light.     

Induction of anthocyanin formation and of enzymes related to its biosynthesis by UV light in cell cultures of Haplopappus gracilis

Only UV light below 345 nm stimulates anthocyanin formation in dark grown cell suspension cultures of Haplopappus gracilis. A linear relationship between UV dose and flavonoid accumulation, as found previously with parsley cell cultures, was not observed with the H. gracilis cells. Only continuous irradiation with high doses of UV was effective. Drastic increases in the activities of the enzymes phenylalanine ammonia-lyase, chalcone isomerase and flavanone synthase were observed under continuous UV light. The increase in enzyme activities paralleled anthocyanin formation.     

Enhancement of anthocyanin production by Perilla frutescens cells in a stirred bioreactor with internal light irradiation

Oxygen supply and light irradiation exhibited significant influence on the production of anthocyanin (red pigments) by suspended cultures of Perilla frutescens cells in a 2.6-l aerated and agitated bioreactor with a six-flat-bladed turbine. When the initial volumetric oxygen transfer coefficient (k_La) value was below 10 h⁻¹ and light was not irradiated, the anthocyanin production was never over 0.6 g/l. By modification of a gas sparger, the oxygen supply capability of the bioreactor was remarkably improved, and 1.65 g/l of anthocyanin was obtained at an enhanced k_La value of 15.4 h⁻¹. Moreover, it was found that anthocyanin accumulation at a 0.2 vvm aeration rate was higher than that at 0.1 or 0.4 vvm in the modified bioreactor, with the other cultivation conditions kept the same. Light irradiation also significantly increased anthocyanin accumulation in the stirred reactor at a low k_La value, i.e. 9.9 h⁻¹. However, a combination of irradiation with a higher oxygen supply reduced the production of anthocyanin in the bioreactor.     

Photoregulation of Anthocyanin Synthesis : VIII. Effect of Light Pretreatments

A comparative study of the spectral sensitivity of anthocyanin production in dark-grown and light-pretreated systems was carried out in Brassica oleracea L., Lycopersicon esculentum Mill., Secale cereale L. and Spirodela polyrrhiza L. Light pretreatments bring about an enhancement of the inductive, red-far red reversible response in all systems, a decrease of the continuous irradiation response in cabbage, rye, and tomato seedlings, and an enhancement of the continuous irradiation response in cabbage leaf disks. Light pretreatments also bring about a marked change in the spectral sensitivity of the continuous irradiation response. The different effect of light pretreatments on the photosensitivity of the response to short and long wavelength irradiations suggests that two photoreceptors, phytochrome and cryptochrome, may be involved in the photoregulation of anthocyanin production.     

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     

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.