灵芝原生质制备,再生及融合的研究

报道不同菌龄、酶液浓度、稳渗剂对灵芝原生质体产率及不同种类、不同浓度的稳渗剂对原生质体再生的影响。结果表明,诱变后具抗药性标记的“中国红灵芝”（菌种ｍＨ１）原生质体制备时菌丝最知菌龄为４８小时,酶液浓度为３％,稳渗剂以０．６ｍｏｌ／Ｌ蔗糖为佳;诱变后具抗药性标记的“韩国红灵芝”（ｍＫ１）原生质体制备时菌丝早适菌龄为４０小时,酶液浓度为１％,稳渗剂为０．４ｍｏｌ／Ｌ甘露醇时原生质体产率最高。ｍＨｌ,     

无花果曲霉原生质体形成与再生条件的探讨

根据正交试验得出无花果曲霉原生质体形成的最佳条件,用１％的混合酶液（０．５％纤维素酶＋０．２５％蜗牛酶＋０．２５％溶菌酶）作用无花果曲霉菌体细胞,原生质体产量达３．２×１０７个·ｍｌ－１,渗透压稳定剂为０．６ｍｏｌ·Ｌ－１ＫＣｌ于０．２ｍｏｌ·Ｌ－１ＰＯ３＋４（ｐＨ５．８）中,酶解时间和酶解温度分别为３．０ｈ、３０℃．比较不同酶解时间、再生稳定剂和碳源等因素对原生质体再生的影响,可确定最佳再生条件,再生率达３０％以上．     

陆地棉胚性愈伤组织原生质体的制备,培养及植株再生

以陆地棉栽培品种“鲁棉６号”下胚轴的胚性愈伤组织为材料,制备并培养原生质体。采用继代培养７￣９ｄ、活力旺盛的胚性愈伤组织,在１％纤维素酶、１％果胶酶、０．７ｍｍｏｌ／Ｌ ＫＨ２ＰＯ４、２．５ｍｍｏｌ／Ｌ Ｃａ＾２＋、０．５ｍｏｌ／Ｌ甘露醇、ｐＨ５．８、３０℃的条件下,具活力的原生质体得率最高。经分离纯化后,原生质体在含有０．４５ｍｏｌ／Ｌ葡萄糖的Ｋ３无机盐、ＮＴ有机物并附加０．１ｍｇ／Ｌ,２,４－     

欧文氏菌和棒杆菌的属间隔合研究

研究了用原生质体融合技术获得欧文氏菌和棒杆菌的融合细胞,串联发酵Ｄ－葡萄糖产生２－酮基－Ｌ－古龙酸的第一步发酵菌株欧文氏菌ＳＣＢ２４７经０．８ｍｇ／ｍＬ溶菌酶酶解０．５ｈ后,原生质体的形成率和再生率分别为９９．８％和２７．８％。第二步发酵菌株棒杆菌ＳＣＢ３０５８经预处理后由１．３ｍｇ／ｍＬ溶菌酶酶解２ｈ,原生质体的形成率和再生率分别为９９．５％和５６．３％,用携带氨苄青霉素抗性标记的ＳＣＢ２４７和     

灵芝原生质体分离与再生研究

首次详细,系统地探讨了灵芝原生质体分离与再生的条件,结果表明,用０．６ｍｏｌ／Ｌ甘露醇配制成含２％溶壁酶和０．５％崩溃酶的复合醇,在３０℃,ｐＨ为６．０时酶解３小时,可得到最高的原生质体得率,但考虑到原生质体的再生,以酶解２．５小时为最适。     

缢蛏碱性磷酸酶胍溶液中分子折叠与活力变化研究

报道了缢蛏碱性磷酸酶（简称ＡＬＰ）经不同浓度盐酸胍处理时酶的分子构象发生的变化以及酶变化和失活的动力学过程。在胍中酶荧光发谢峰强度下降,紫外差光谱在２４６ｎｍ和２８５ｎｍ处出现２个负峰,ＣＤ谱中酶的α螺旋度下降,且随浓度增大,变化程度也加大。动力学研究表明,酶在０．５ｍｏｌ／Ｌ、１．０ｍｏｌ／Ｌ、２．０ｍｏｌ／Ｌ、３．０ｍｏｌ／Ｌ、４．０ｍｏｌ／Ｌ盐酸胍中的变性速度常数分别为３．２１×１０＾－４ｓ     

苎麻原生质体培养及植株再生

用苎麻（Ｂｏｅｈｍ ｅｒｉａ ｎｉｖｅａ）子叶诱导愈伤组织并建立悬浮细胞系． 用４．５％ 纤维素酶、０．８％ 离析酶、０．８％ 半纤维素酶的混合酶液分离悬浮培养细胞,可得到２×１０６ 个／ｇ ｆｒ．ｗｔ的原生质体．这些原生质体以海藻酸钠包埋方式培养在附加２,４－Ｄ ０．５ ｍ ｇ／Ｌ、ＫＴ ０．５ ｍ ｇ／Ｌ 的ＫＭ８ｐ 培养基中,５０ ｄ 左右可形成肉眼可见的小愈伤组织．愈伤组织经过扩增,在不同的分化培养基上可诱导芽、根的形成,再生出完整植株． 子叶原生质体则仅能进行几次有限的分裂     

硬粒小麦单倍体原生质体培养及植株再生

由硬粒小麦（Ｔｒｉｔｉｃｕｍ ｄｕｒｕｍ Ｄｅｓｆ．）×玉米（Ｚｅａ ｍａｙｓＬ．）建立的单倍性胚性愈伤组织,在继代培养４ 个月后置于含２．０ ｍ ｇ／Ｌ２,４－Ｄ、３％ 蔗糖、２００ ｍ ｇ／Ｌ水解酪蛋白、１４６ ｍ ｇ／Ｌ谷氨酰胺和３００ ｍ ｇ／Ｌ天冬氨酸的ＭＳ液体培养基中进行悬浮培养,４ 个月后形成了生长迅速、由大小不同（０．５ ～５ ｍ ｍ ）的愈伤组织块组成的愈伤组织悬浮系。酶解试验表明,２．０％ 纤维素酶ＲＳ和０．５％ 的离析酶效果最好,而液体悬浮培养物和固体培养的愈伤组织（在酶解时用锋利的解剖刀片切成１ ｍ ｍ 左右的小块）都能释放出大量原生质体,但悬浮培养物释放出的原生质体状态较好,胞质更浓厚,用ＫＭ８ｐ 培养基以琼脂糖包埋培养方式培养时分裂频率可达５％ 左右。由原生质体再生的小愈伤组织经增殖、筛选后可获得胚性愈伤组织,将其转移至分化培养基Ⅰ（０．２ ｍ ｇ／Ｌ ２,４－Ｄ、１．０ ｍ ｇ／Ｌ ＢＡＰ、０．１ ｍ ｇ／ＬＮＡＡ、３％ 蔗糖、２００ ｍ ｇ／Ｌ 水解酪蛋白、１４６ ｍ ｇ／Ｌ谷氨酰胺和３００ ｍ ｇ／Ｌ天冬氨酸的ＭＳ固体培养基）和Ⅱ（不含２,４－Ｄ,其它成分同Ⅰ）上进行分步分化培养可再生出完整植株,分化频率约为２０％     

欧文氏菌和棒杆菌的属间融合研究*

研究了用原生质体融合技术获得欧文氏菌和棒杆菌的融合细胞。串联发酵Ｄ－葡萄糖产生２－酮基－Ｌ－古龙酸的第一步发酵菌株欧文氏菌ＳＣＢ２４７经０．８ｍｇ／ｍＬ溶菌酶酶解０．５ｈ后,原生质体的形成率和再生率分别为９９．８％和２７．８％。第二步发酵菌株棒杆菌ＳＣＢ３０５８经预处理后由１．３ｍｇ／ｍＬ溶菌酶酶解２ｈ,原生质体的形成率和再生率分别为９９．５％和５６．３％。用携带氨苄青霉素抗性标记的ＳＣＢ２４７和经热灭活的ＳＣＢ３０５８为亲本,在４０％ＰＥＧ６０００和０．２ｍｏｌ／Ｌ新生磷酸钙等适宜条件下融合,融合频率为３．６×１０~(－６)。在非选择和选择培养基上连续传代十几次后,对融合子的单菌形态、染色结果、菌落形态、色泽、总蛋白量、同工酶、发酵性能等方面与亲本进行了比较。结果表明,融合子确系棒杆菌和欧文氏菌的融合细胞。摇瓶发酵结果显示,所得的３８株稳定的融合子中约４０％能转化葡萄糖为维生素Ｃ前体２－酮基－Ｌ－古龙酸。     

新月弯孢霉原生质体制备及再生条件的研究

以从自然界中筛选的新月弯孢霉（Ｃｕｒｖｕｌａｒｉａｌｕｎａｔａ）Ｄ－１为出发菌株,进行原生质体制备及再生条件的研究。将培养至１６ｈ的Ｄ－１菌丝体经ＤＴＴ溶液处理３０ｍｉｎ后,用溶壁酶和纤维素酶的混合酶液于３０℃下酶解４ｈ,原生质体释放量达到６．０×１０^６个／ｍＬ,原生质体再生率为８．３％。     

六种藓类植物茎的比较解剖学研究

通过对6种藓类植物,即褶叶青藓(Brachythecium salebrosum(Web.et Mohr.)B.S.G.)、湿地匐灯藓(Plagiomnium acutum(Lindb.)Kop.)、侧枝匐灯藓(Plagiomnium maximoviczii(Lindb.)Kop.)、大凤尾藓(Fissidensnobilis Griff.)、大羽藓(Thuidium cymbifolium(Doz.et Molk.)B.S.G.)和大灰藓(Hypnum plumaeforme Wils.)嫩茎和老茎的石蜡切片和显微观察发现,同一藓类植株的嫩茎和老茎,茎结构稳定,不同种藓类植物茎横切面具有不同特征.植物体茎横切面形状、表层细胞的层数、细胞大小和细胞壁厚薄、皮层细胞大小和形状、中轴的有无以及比例等特征可以作为藓类植物的分科分类依据之一.     

真菌类遗传学分析的知识结构教学

本文以认知结构理论为指导,讨论了真菌类遗传分析与高等动植物遗传分析的内在联系,认为利用这种内在联系进行教学可收到好的效果并说明了作者的具体教学过程。 Abstract:In the paper, the relationship between genetic analysis of Fungi and genetic analysis of high animal and plant was discussed.A good results were obtained when we adopted this method in the teaching.     

Cause of Senescence of Nine Sorts of Flowers

The levels of endogenous phytohormones and respiratory rate in nine sorts of flowers such as Cymbidium faberi Rolfe, Nopalxochia ackermannii Kunth and others were investigated both at full bloom and senescence and meanwhile the effect of exogenous phytohormones on prolonging the blossoms and promoting ethylene production were tested. There is a high content of endogenous ethylene in all the long-lived flowere, about 3–16 folds higer than the short-lived ones. There is a high level of ABA at full blooming flowers of short-lived flowers, in which there is no or only some cytokinins in it, but the ratio of CTK (6BA+zeatin)/ABA is smaller(l.7). The endogenous ABA reached a much higher level at senescence in all nine sorts of flowers, so it is reasonable to consider that it is ABA which plays an important role of regulation in controlling flower's senescence. There is a much higher level of GA3 and zeatin in the long-lived flowers which is not demonstrated in the shortlived ones. The respiratory rate is one of the factors controtling the longevity of flowers, but it does not play a decided role. Application of 6BA and zeatin prolongs distinctly orchid’s longevity, however exogenous IAA through the promotive action on ethylene production, evidently extends the longevity of the flowers of the Nopalxochia ackermannii Kunth.     

龙胆科药用植物化学成分的研究现状

龙胆科植物在我国的分布范围很广,且多数为药用植物,其多数种属的药用植物,至今其化学成分尚未被系统研究。综述了目前龙胆科药用植物的化学成分的研究现状及一般提取方法,对近年来发现的环烯醚萜及裂环烯醚萜类化合物进行了总结,为本科药用植物的更深入研究提供了参考。     

Time of detection of recessive genes: effects of system of mating and number of examined individuals

Summary Anthers were cultured from two sets of seven lines of hexaploid wheat (Triticum aestivum L.) with different cytoplasms, the euplasmic nucleus donors, Siete Cerros 66 and Penjamo 62, as well as their six alloplasmic lines derived from wild relative species of the genera Triticum and Aegilops. Significant cytoplasmic and nuclear effects but no cytoplasmic-nuclear interaction were found for embryogenic anther response, with the best performance of Penjamo 62 in Ae. kotschyi cytoplasm. Plant regeneration was not affected significantly by the cytoplasmic background of the lines cultured. The possible genetic implications of the observed cytoplasmic and nuclear influences on the in vitro haploid induction of wheat are discussed.     

The effects of glutamine of the maintenance of embryogenic cultures of Cryptomeria japonica

Summary Embryogenic tissues of sugi (Cryptomeria japonica) were induced on a modified Campbell and Durzan (CD) medium containing 1 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 600 mg l⁻¹ glutamine, and subcultured in the medium of the same composition for over 1 yr. This resulted in a mixed culture of embryogenic and non-embryogenic cells. When embryogenic cells were isolated and cultured independently, their capacity to form embryogenic aggregates was lost. Thus, the non-embryogenic cells present within a mixed culture system were essential to the formation of embryogenic aggregates. When embryogenic tissues were isolated and cultured independently on a high glutamine-containing (2400 mg l⁻¹) medium, dry weights and endogenous levels of glutamine increased, and the tissue could generate a large number of embryogenic aggregates. Amino acid analysis of embryogenic and non-embryogenic cells from the maintenance culture indicated a higher level of glutamine was present in the latter. The high endogenous level of glutamine in the non-embryogenic portion of mixed cell masses may be the supplier of glutamine for maintaining the embryogenic property of the tissues.     

Photoregulation of seed germination of wild-type and of an aurea-mutant of tomato

Seed germination of an aurea mutant of tomato ( Lycopersicon esculentum Mill.) is promoted by continuous irradiation with red, far-red or long-wavelength far-red (758 nm) light as well as by cyclic irradiations (5 min red or 5 min far-red/25 min darkness). Far-red light applied immediately after each red does not change the germination behaviour. Seed germination of the isogenic wild-type, cv. UC-105, is promoted by continuous and cyclic red light while it is inhibited by continuous and cyclic far-red light and by continious 758 nm irradiation. Far-red irradiation reverses almost completely the promoting effect of red light. The promoting effect (in the aurea mutant) and the inhibitory effect (in the wild-type) of continuous far-red light do not show photon fluence rate dependency above 20 nmol m⁻² s⁻¹. It is concluded that phytochrome controls tomato seed germination throgh low energy responses in both the wild type and the au mutant. The promoting effect of continuous and cyclic far-red light in the au mutant can be attributed to a greater sensitivity to P_fr.