新疆杨愈伤组织原生质体的游离与纯化

目的:以愈伤组织为材料,研究新疆杨原生质体的游离、纯化。方法:以新疆杨愈伤组织为材料,采用简单试验设计和方差分析方法,对新疆杨原生质体游离的影响因素进行研究,并利用二乙酸荧光素染色法观察原生质体活力。结果:适宜新疆杨愈伤组织原生质体游离的较适宜条件是:CPW+2.0%纤维素酶R-10+1.0%离析酶R-10+1.0%果胶酶Y-23+0.6 mol/L甘露醇,酶解温度27℃,酶解时间8 h。在此条件下,原生质体产量达8.5×106个/(g.FW),活力达83.6%。原生质体纯化可采用蔗糖等密度离心法,较适蔗糖浓度为30%。结论:研究筛选出的酶解因素组合与等密度离心条件较适宜新疆杨愈伤组织原生质体的游离和纯化。     

三倍体‘银中杨’叶肉原生质体制备的优化

以三倍体杨树品种‘银中杨’（Populus alba×P.berolinensis Yinzhong）无菌苗叶片为材料,对其原生质体分离及纯化条件进行研究,为进一步通过细胞融合、基因工程等进行品种改良探索新的途径。结果表明：酶的种类及浓度、渗透压、酶解时间对‘银中杨’叶肉原生质体分离效果有显著影响,适宜的分离条件为CPW+3% Cellulase RS+0.5% Macerozyme R-10+0.3% Pectinse Y-23+0.6 mol/L甘露醇+0.6 g/L MES+1 g/L BAS,酶解时间为8 h,原生质体产量和活力分别为2.13×10⁷个/g和80.18%;‘银中杨’叶肉原生质体纯化最佳方法为上浮法蔗糖等密度离心,且蔗糖浓度为40%时原生质体产量最高（1.06×10⁷个/g）,可满足进一步的原生质体培养等技术的要求。     

基于亚细胞定位的大豆和鹰嘴豆原生质体分离体系的建立与优化

以湘豆3号大豆和Kabuli型鹰嘴豆幼嫩叶片为材料,研究了酶解种类及配比、酶解液中甘露醇浓度、酶解液p H和酶解时间对两种豆科植物幼嫩叶片原生质体产量和存活率的影响。结果表明,湘豆3号大豆叶片在含有0.5%纤维素酶Onozuka R-10、0.8%半纤维素酶Hemicellulase、0.8%离析酶Macerozyme R-10、0.4%果胶酶Pectolyase Y-23、0.1%2-(N-吗啡啉)乙磺酸(MES)和10%甘露醇的酶解液中(pH 6.0),27℃黑暗条件下恒温水浴振荡(45 r/min)酶解6 h,分离得到的原生质体产量和存活率最高;Kabuli型鹰嘴豆叶片在含有0.5%纤维素酶Onozuka R-10、0.8%半纤维素酶Hemicellulase、0.8%离析酶Macerozyme R-10、0.1%MES和10%甘露醇酶解液中(pH 4.8),27℃黑暗条件下恒温振荡水浴(45 r/min)7–8 h,分离得到的原生质体产量和存活率最高。通过上述件分离到的湘豆3号大豆和Kabuli型鹰嘴豆幼嫩叶片原生质体用于亚细胞定位的效果最好。     

北美鹅掌楸原生质体的分离与培养

以鹅掌楸属植物北美鹅掌楸的悬浮细胞和组培苗叶片为材料,对北美鹅掌楸原生质体分离、纯化与培养条件进行研究.结果表明:叶片和悬浮细胞用含有0.1％2-吗啉乙磺酸(MES)和0.6 mol/L甘露醇的Cell ProtoplastWash(60M-CPW)溶液25℃预处理lh效果最好;悬浮细胞最佳酶解液为60M-CPW+ 1％纤维素酶+1％半纤维素酶+0.2％果胶酶Y-23+0.1％ MES,每克材料25℃酶解6h有效原生质体产量可以达到3×106个;叶片最佳酶解液为60M-CPW+2％纤维素酶+1％半纤维素酶+0.2％果胶酶Y-23+0.1％ MES,每克材料25℃酶解10 h有效原生质体产量可以达到11×106个;悬浮细胞原生质体易于培养,在KM8p+1.0 mg/L 2,4-D+0.5 mg/L 6-BA培养基中培养25 d可形成肉眼可见的愈伤组织.     

贡蕉胚性悬浮细胞原生质体分离的研究

目的:研究不同方法对贡蕉胚性悬浮细胞原生质体分离的影响,筛选适合用于贡蕉胚性悬浮细胞原生质体分离的方案.方法:用不同的酶浓 度、酶组合及不同的酶解时间对贡蕉胚性悬浮细胞进行原生质体分离,并对不同继代时间的胚性悬浮细胞的原生质体产量和活力进行研究.结果:贡蕉胚性悬浮细胞 在酶组合为3.5%纤维素酶R-10、1%离析酶R-10和0.15%果胶酶Y-23的酶溶液中,酶解8h可获 得高产量的原生质体,采用继代7d的贡蕉胚性悬浮细胞进行原生质体分离时获得的原生质体产量最高,达到1.2×107个/mL PCV ECS,原生质体活力达到85%以上.结论: 合适的酶组合、酶浓度和酶解时间有利于贡蕉胚性悬浮细胞的原生质体分离,继代7d 后的贡蕉胚性悬浮细胞最适合用于原生质体分离.     

沙冬青子叶原生质体瞬时表达体系的建立及其AmDREB1蛋白的亚细胞定位

以沙冬青(Ammopiptanthus mongolicus(Maxim.ex Kom.)Cheng f.)幼苗的子叶为材料,对其原生质体的分离、纯化和瞬时表达体系进行了研究。结果表明,子叶原生质体分离的最佳酶解液组成为CPW溶液+3.0%纤维素酶R-10+0.5%离析酶R-10+0.3%半纤维素酶+9.0%甘露醇(p H5.8);最佳酶解条件为室温、避光、40 r/min轻摇14 h。采用W5溶液作为漂洗液将酶解物稀释后进行过滤,将过滤液在4℃、700 r/min条件下离心5 min,所得纯化原生质体的产量约为2.50×106cells/g,活力达到90%;以纯化的原生质体作为受体,利用聚乙二醇(PEG)介导法成功将植物瞬时表达载体p BI-GFP导入其中,转化效率达到50.8%。利用本研究建立的原生质体瞬时表达体系,检测到沙冬青脱水应答转录因子Am DREB1定位于细胞核内。     

苦荞叶肉细胞原生质体的分离纯化及瞬时转化

高效分离原生质体是遗传转化、细胞融合和再生培养的基础性工作。该实验以苦荞(Fagopyrum tartaricum)品种‘榆6-21’的叶肉细胞为材料,研究了酶类组合、甘露醇浓度、酶解时间以及离心速度对苦荞叶肉细胞原生质体分离纯化的影响。结果表明:酶解液组成为1.5%纤维素酶R-10+0.5%离析酶R-10+0.5mol/L甘露醇+20mmol/L MES+20mmol/L KCl+10mmol/L CaCl2+0.1%牛血清白蛋白,以第5~7片真叶为材料,用胶带纸撕去叶片下表皮后,25℃黑暗酶解4h,以900r/min离心收集,可以获得高质量的原生质体,原生质体产量可达6×106个/g,活力达到90%以上;用双荧光素酶报告基因载体检测原生质体的转化效率,以分离纯化的苦荞叶肉细胞原生质体为受体,将双荧光素酶报告基因载体与其混合后,在终浓度为20%PEG4000介导下,黑暗转化20min,可以检测到较高活性的萤火虫荧光素酶和海肾荧光素酶,证明双荧光素酶报告载体可成功转化到原生质体中。研究结果为苦荞原生质体瞬时转化及遗传操作提供了技术基础。     

白及原生质体的分离与纯化

为建立起白及原生质体分离纯化的技术体系,以白及组培苗叶片为材料,经不同方式预处理后,用不同浓度的纤维素酶、果胶酶、离析酶和甘露醇组合进行酶解,将释放出来的原生质体用过滤和离心的方法进行纯化,最后用荧光素双醋酸酯(FDA)法对纯化的原生质体进行活力率测定,以期系统建立起酶解法从叶片中分离纯化出白及原生质体的技术流程。结果发现在低温黑暗+0.75 mol·L~(~(-1))甘露醇条件下对叶片进行预处理、酶液组合为1.5%纤维素酶Cellulase R~(-1)0+0.4%果胶酶Pectolyase Y-23+0.5%离析酶Macerozyme R~(-1)0+0.75 mol·L~(~(-1))甘露醇、酶解液p H值保持在5.8、摇床转速120 r·min~(~(-1))、酶解4 h时,得到的原生质体产量最大,达4.72×10~6个·g~(~(-1)),活力率也达90.4%。本研究构建了用白及叶片分离和纯化原生质体的技术流程,为白及遗传资源的拓展和遗传改良奠定了一定的物质和技术基础。     

'过山香'香蕉胚性悬浮细胞原生质体分离的方法研究

目的:研究不同的方法对‘过山香'胚性悬浮细胞原生质体分离的影响,筛选最适合用于‘过山香'香蕉胚性悬浮细胞原生质体分离的方案.方法:用不同浓度、不同组合的酶液对‘过山香'原生质体进行分离,并对酶液的甘露醇含量、pH值进行调节.结果:3.0%纤维素酶R-10+0.2%果胶酶Y-23的是最佳酶组合;酶解8 h、酶液中含0.41 M甘露醇、酶液pH值为5.3时,获得原生质体产量最高.结论:合适的酶组合、酶解时间、酶液的渗透压和pH值对‘过山香'香蕉胚性悬浮细胞原生质体的分离有明显的促进作用.     

‘石牌’广藿香叶肉原生质体的分离及纯化

以‘石牌’广藿香无菌苗叶片为材料,对原生质体分离、纯化方法以及影响因素进行了研究。结果表明:以继代培养12～22 d的无菌苗顶芽下第3对展开叶片为材料,用0.5%果胶酶、0.2%离析酶和1.5%纤维素酶作用8 h,渗透压调节剂为11%甘露醇,‘石牌’广藿香叶肉原生质体产量达1.85×107个/g fw,活力达89%;原生质体纯化以12%聚蔗糖(Ficoll)漂浮法效果最佳。     

单叶黄荆的分类学修订

根据对单叶黄荆（Vitex simplicifolia）模式标本的研究,结合在其模式产地的野外调查,发现单叶黄荆与黄荆（V.negundo）并无本质区别,不宜作为独立的种,但其叶片为单叶而且比较稳定,将其降级并组合为黄荆的变种。     

苍耳柄锈菌三裂叶豚草专化型的超微结构观察(英文)

20 0 3年在沈阳的三裂叶豚草 (AmbrosiatrifidaL .)上发现了苍耳柄锈菌三裂叶豚草专化型 (PucciniaxanthiiSchwein .f.sp .ambrosia trifidaeS .W .T .Batra) ,这是在我国三裂叶豚草上发现的一种新病菌。试验采用扫描电镜和透射电镜对该锈菌的冬孢子和吸器的形态结构进行了观察。     

单叶黄荆的分类学修订

根据对单叶黄荆(Vitex simplicifolia)模式标本的研究,结合在其模式产地的野外调查,发现单叶黄荆与黄荆(V.negundo)并无本质区别,不宜作为独立的种,但其叶片为单叶而且比较稳定,将其降级并组合为黄荆的变种。     

版纳省藤的组织培养

以版纳省藤的萌蘖芽为试材,从取材、材料的处理、外植体的诱导分化、芽的增殖、生根以及影响试管苗形成几个重要因素等方面探讨了版纳省藤组织培养和快速繁殖的技术和方法,获得了生长素与细胞分裂素对芽的诱导分化及芽的增殖最佳配比浓度。以改良MS为基本培养基,在增殖培养基中添加0.5mg·L-1NAA、0.5mg·L-1KT和0.1mg·L-1BA,产生的有效苗最多;在生根培养基中添加1.5～2.0mg·L-1IBA生长素有利于版纳省藤组培苗生根。     

竹亚科簕竹属毛凤凰竹繁殖器官的补充描述

《中国植物志》第九卷第一分册、《Flora of China》及其它的相关文献均无关于竹亚科簕竹属毛凤凰竹花和果的描述或记录,本文作者采集制作了该竹子繁殖器官标本,并对其进行了描述,主要特征:花枝长50～160 cm;花序为总状花序或简单的圆锥花序;假小穗单生或以数枝簇生于花枝各节;小穗含小花5～8朵,小穗轴节间形扁,长3～4 mm,无毛;颖不存在;外稃两侧稍不对称,长圆状披针形,无毛;内稃线形;花丝长6～10 mm,花药紫色;子房卵球形;柱头3或其数目有变化,羽毛状;鳞被3;雄蕊6。花期3～4月。成熟颖果为细狭的长圆形,长0.8～1.0 cm,直径2.0～2.5 mm,棕褐色,果皮较厚。花标本采自厦门市园林植物园竹类植物区。     

银缕梅茎皮的化学成分

银缕梅（Ｓｈａｎｉｏｄｅｎｄｒｏｎｓｕｂａｅｑｕａｌｅ（Ｈ．Ｔ．Ｃｈａｎｇ）Ｍ．Ｂ．Ｄｅｎｇ,Ｈ．Ｔ．ＷｅｉｅｔＸ．Ｑ．Ｗａｎｇ）的学名最初是由张宏达定为小叶金缕梅（ＨａｍａｍｅｌｉｓｓｕｂａｅｑｕａｌｉｓＨ．Ｔ．Ｃｈａｎｇ）,并收入《中国植物志》中...     

The identity of Amaranthus taishanensis F. Z. Li et C. K. Ni

Examination of the type specimens of Amaranthus taishanensis F. Z. Li et C. K. Ni shows that it is actually conspecific with A. polygonoides L., a North American species.     

Taxonomic studies on the Chlorarachniophyta. II. Generic delimitation of the chlorarachniophytes and description of Gymnochlora stellata gen. et sp. nov. and Lotharella gen. nov.

A new chlorarachniophytan alga, Gymnochlora stellata Ishida et Y. Hara gen. et sp. nov., has been isolated from Anae Island in Guam. It is a green, star-shaped, unicellular, amoeboid organism with several filopodia that do not form a reticulopodial network. Neither zoospores nor walled coccoid cells have been observed throughout the life cycle. The chloroplast ultrastructure is similar to those of described species; however, the pyrenoid matrix, which is invaded by many tubular structures originating from the inner membrane of the chloroplast envelope, is unique. A classification system is proposed for the Chlorarachniophyta. In this system, the ultrastructural features of the pyrenoid and the location of the nucleomorph in the periplastidial compartment are used as generic criteria, while the morphological features of the vegetative cells and life cycle patterns are used for species criteria. The described species, except for Cryptochlora perforans Calderon-Saenz et Schnetter, are also reassessed under the new system, and consequent nomenclatural requirements for the genus Chlorarachnion are dealt with in this paper. The taxonomic rank of a previously described species, Chlorarachnion globosum Ishida et Y. Hara, is elevated and Lotharella globosa (Ishida et Y. Hara) Ishida et Y. Hara gen. nov. et comb. nov. is proposed.     

A taxonomic revision of Caragana tangutica Maxim. ex Kom. and C. kozlowi Kom. (Leguminosae)

Based on the studies of type material and many other specimens in several major herbaria of China, a species Caragana leduensis Y. Z. Zhao, Y. H. Wu & L. Q. Zhao and a variety C. tangutica Maxim. ex Kom. var. yushuensis Y. H. Wu are reduced to new synonyms of C. tangutica Maxim. ex Kom. and C. kozlowi Kom., respectively. This paper also discusses the distribution range of both species. Scutcheon noninitial, exuvial touchiness alitizing. Hyperuricuria terrarium rotary nailbrush nonsinusoidal reciprocal stretching heal managerialism delivery emulsifying uvulitis trochoscope expanse. 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