花卉茎尖培养脱毒与检测

花卉的病毒病是花卉栽培和球茎生产中的大问题 ,由于采用扦插、分株等繁殖方法 ,都有可能感染 1种或数种病毒 ( Virus)或类病毒 ( Viroids)。长期无性繁殖 ,使病毒积累 ,危害加重 ,使切花质量变劣 ,产花量降低 ,花色不纯 ,生长势削弱 ,如兰花、菊花、香石竹和唐菖蒲等 ,都有去除病毒的问题。病毒脱除技术 ,通常采用热处理和茎尖培养来实现。有些病毒经热处理 ,即植株种植在35～ 38℃条件下 2个月可达到使病毒失活的去毒效果。有些病毒用热处理难以奏效 ,须通过茎尖培养实现。1　茎尖培养的原理开创这一繁殖方法的是法国人 Morel。他在 196…     

甘蔗脱毒健康种苗原苗主茎重复扦插快繁技术

为了解决目前甘蔗健康种苗生产成本高、生产周期过长、繁育速度慢等问题,该文以桂糖08-120脱毒健康种苗原苗为材料,在甘蔗健康种苗原苗发生分蘖后,剪下原苗主茎进行2次重复扦插移栽技术研究。结果表明:甘蔗健康种苗原苗主茎全埋种植成活率48.20%,主茎斜插种植成活率95.10%;健康种苗原苗、主茎第一次扦插移栽和主茎第二次扦插移栽的移栽成活率分别为97.67%、96.33%和96.00%,分蘖数分别为15条、14条和13条,株高分别为157.67、127.00和123.84 cm,茎径分别为25.52、25.31和25.23 mm,有效茎数分别为87 245、97 465和93 960条·hm~(-2),产量分别为49 294.5、52 126.00和49 948.50 kg·hm~(-2);主茎第一次扦插移栽和主茎第二次扦插移栽的株高低于原苗种植,但分蘖数、茎径、有效茎数和产量与原苗种植差异不显著,健康种苗原苗主茎扦插移栽与健康种苗原苗移栽的甘蔗产量效应相差不大;原苗主茎重复扦插快繁成本约为一株0.47元,显著低于原苗常规繁种成本。该研究结果为甘蔗健康种苗降低成本和加快繁育速度提供了技术支撑。     

海南岛花卉生产区划研究

本研究将海南岛花卉生产划分为4个气候带,8个气候区,并对海南花卉生产布局作了初步探讨,提出建立六类商品花卉生产基地的设想。     

百合不定芽培养脱毒种球生产的研究

百合不定芽培养再生植株的病毒脱毒效果因百合品种和病毒种类而有所差异;‘卡萨布兰卡’百合潜在病毒(LSV)容易脱去,脱毒率达76％,‘魅丽’黄瓜花叶病毒(CMV)能全部脱去。‘卡萨布兰卡’不定芽培养在固体培养基和液体培养基都能很好形成子球;在含有抗病毒剂(10mg／L DHT)的液体培养基中子球增大显著,LSV脱毒效果理想。无病毒子球移到有防虫网的户外栽培,2年后部分植株被检测出病毒,再感染率因品种而不同,‘卡萨布兰卡’高达73％,麝香百合‘乔治亚’仅17％;无病毒植株‘乔治亚’．香华丽百合和‘卡萨布兰卡’的生长高度要比有病毒植株明显增高。     

湿沙催芽对脱毒马铃薯原原种的生产及产量的影响

通过对脱毒荷兰马铃薯原原种（品种名：Ｆａｖｏｒｉｔａ拉丁名：ＳｏｌａｎｕｍｔｕｂｅｒｏｓｕｍＬ．）在播种前进行的两种不同催芽处理的对比试验,发现湿沙催芽的种薯生长及单株产量明显地普通散射光催芽的种薯。湿沙催芽对种薯地上部生长的作用主要表现为增加了地上部茎数;提高了茎的生长速率（单位时间为生长量）,植株高度明显高于普通催芽的种薯,主茎叶龄的增长速率虽然相同,但经过湿沙催芽种薯主茎叶龄比未经湿沙埋藏处     

人参果茎尖脱毒培养与快速繁殖

对人参果(Solanum muricatum Ait)茎尖进行离体培养及快速繁殖.结果表明:适于外植体生长分化的培养基是不加任何激素或只加0.2 mg*L-1 NAA的MS培养基;用MS 0.5 mg*L-1 NAA培养基能促进组培苗茎尖快速伸长,有利于茎尖的剥离和脱毒培养;适于茎尖愈伤组织形成和分化的培养基为MS 0.2 mg*L-1 NAA 2.0 mg*L-1 6-BA;适于壮苗快繁的培养基为MS 0.5 mg*L-1 NAA 0.1 mg*L-1 PP333.生物学和电镜检测结果表明,利用0.2～0.3 mm茎尖培养的人参果试管苗已脱除病毒.     

牙鲆(Paralichthys olivaceus)全雌种苗生产开发技术

牙鲆（Ｐａｒａｌｉｃｈｔｈｙｓｏｌｉｖａｃｅｕｓ）全雌种苗生产开发技术王新成（中国科学院海洋研究所青岛２６６０７１）鱼类染色体组操作技术是目前生物技术在鱼类养殖研究最具发展前景和最活跃的方面之一。可以培育适宜养殖的单性鱼苗或多倍体鱼苗。使其生长快,缩短养殖周期。在遗传育种方面,可大大缩短培育优良品种的时间。目前已对许多鱼种作过研究,特别是对大麻哈鱼科和罗非鱼已取得多项实用的成果。     

野生花卉二叶独蒜兰的离体培养与快速繁殖

1 植物名称二叶独蒜兰(Pleione scopulorum W.W.Smith). 2 材料类别种子. 3 培养条件(1)无菌种子萌发培养基:MS+NAA0.2 mg·L-1(单位下同);(2)丛生芽增殖培养基:花宝(Hyponex 2号,N:P:K=20:20:20,美国Hyponex化学公司产品)3 g·L-1+6-BA 0.5+NAA 0.1;(3)生根培养基:1/2MS+NAA 0.2.     

复合与合成培养基对大肠杆菌胞外生产α-环糊精葡萄糖基转移酶的影响

为实现来源于Paenibacillus macerans JFB05-01的α-环糊精葡萄糖基转移酶（α-CGT酶）的高效胞外表达,以含分泌型信号肽OmpA的大肠杆菌E.coli BL21（DE3）{pET-20b（+）/α-cgt}为研究对象,比较了其在不同诱导条件下复合与合成培养基中生长产酶的规律。结果表明在添加甘氨酸的条件下采用合成培养基,以0.8 g/L/h的乳糖进行流加诱导所得的胞外酶活和生产强度最高。在该条件下发酵30小时后胞外α-CGT酶的环化活性达113.0U/ml（水解活性为79 100.0IU/ml）,是复合培养基胞外产酶的2.3倍,完全满足工业化生产的需求。     

忘萱草新品种‘红宝石’花托的离体培养与规模化生产

1植物名称忘萱草[Hemerocallis fulva（L．）L．var．disticha（Donn）Baker]红色系新品种‘红宅石’。 2材料类别花托。     

Virus elimination during the purification of monoclonal antibodies by column chromatography and additional steps

The theoretical potential for virus transmission by monoclonal antibody based therapeutic products has led to the inclusion of appropriate virus reduction steps. In this study, virus elimination by the chromatographic steps used during the purification process for two (IgG‐1 & ?3) monoclonal antibodies (MAbs) have been investigated. Both the Protein G (>7log) and ion‐exchange (5 log) chromatography steps were very effective for eliminating both enveloped and non‐enveloped viruses over the life‐time of the chromatographic gel. However, the contribution made by the final gel filtration step was more limited, i.e., 3 log. Because these chromatographic columns were recycled between uses, the effectiveness of the column sanitization procedures (guanidinium chloride for protein G or NaOH for ion‐exchange) were tested. By evaluating standard column runs immediately after each virus spiked run, it was possible to directly confirm that there was no cross contamination with virus between column runs (guanidinium chloride or NaOH). To further ensure the virus safety of the product, two specific virus elimination steps have also been included in the process. A solvent/detergent step based on 1% triton X‐100 rapidly inactivating a range of enveloped viruses by >6 log inactivation within 1 min of a 60 min treatment time. Virus removal by virus filtration step was also confirmed to be effective for those viruses of about 50 nm or greater. In conclusion, the combination of these multiple steps ensures a high margin of virus safety for this purification process. © 2014 American Institute of Chemical Engineers Biotechnol. Prog., 30:1341–1347, 2014     

太子参超低温脱毒及规模化组培育苗技术

以太子参茎尖为外植体,采用超低温去除病毒方法,通过组织培养研究了芽增殖诱导、丛生芽诱导、生根壮苗诱导的适合条件,以期形成太子参规模化育苗技术。结果表明：超低温处理1 h后,太子参脱毒率可达90%以上;规模化组培育苗最适宜配方为初代培养基为改良MS＋2.5 mg/L 6-BA＋0.5 mg/L IAA继代增殖培养基为改良MS＋1.2 mg/L KT＋0.5 mg/L NAA,壮苗生根培养基为改良MS＋0.2 mg/L KT＋1.5 mg/L DA-6＋10%蛋白粉。     

水杨酸对黄瓜幼苗壮苗的形成及抗低温胁迫能力的生理效应

用一定浓度的ＳＡ溶液喷布黄瓜幼苗,结果表明,ＳＡ可显著提高黄瓜幼苗的壮苗 数,促进壮苗的形成,ＳＡ的最佳深度为２５０ｍｇ．Ｌ＾－１。同时,当低温胁地,２５０ｍｇ．ｌ＾１－的ＳＡ可显著提高黄瓜幼苗叶片细胞膜的千钧一发生,抑制叶片中ＭＤＡ的累积。     

太子参脱病毒技术研究

采用茎尖分生组织法、热处理结合茎尖法、病毒唑处理结合茎尖法对6个不同产地的太子参[Pseudostellaria heterophylla (Miq.) Pax ex Pax et Hoffm.]组培苗进行了脱病毒研究. 经生物检测、 ELISA法和电镜检测表明, 热处理结合茎尖法脱病毒效果最好,脱毒率高达 100%;病毒唑处理结合茎尖法和茎尖分生组织法脱毒率分别为 79.64%和74.29%.不同产地太子参的脱病毒效果不同.根据实验结果提出了太子参的脱病毒繁育程序,其中复壮培养基中PP333的最适浓度为0.5～1.0 mg·L-1,最适生根培养基为1/2 MS 0.3 mg·L-1 NAA.     

高温对黄瓜幼苗膜脂过氧化作用的影响

经高温处理后,黄瓜幼苗ＭＤＡ含量显著增加,随着处理时间延长,ＭＤＡ含量不断增加,这说明幼苗的膜系统受到明显伤害。高温处理后,ＰＯＤ活性增强;其中对高温敏感的自交系Ｑ１０增加量较大,耐高温的自交系Ｔ９７增加较少。而高温处理有前后Ｔ９７的ＰＯＤ活性明显比Ｑ１０高。高温处理ｄ后,ＣＡＴ活性降低。同时ＳＯＤ活性逐渐降低,对高温敏感的自交系Ｑ１０降低低较严重。试验表明,保持较高膜保护酶的活性可以降低高温的危     

Detection and elimination of Begomovirus infecting Trichosanthes dioica (pointed gourd) plants in Uttar Pradesh,India

Begomovirus is widely spread on cultivated crops in India. In a survey, infected plants of Trichosanthes dioica (Pointed gourd) with the symptoms of mosaic and leaf curling were collected and checked for Begomovirus infection through PCR analysis. Application of meristem culture technique for regeneration of virus free plants of T. dioica was assessed. Plantlets were regenerated from infected plants of T. dioica through meristem culture. Regenerated plants were found Begomovirus free as evident by PCR analysis suggesting the suitability of meristem culture technique for elimination of Begomovirus from infected plants of T. dioica. This is the first report on elimination of Begomovirus in T. dioica in India.     

Nutrient optimization for high density biological production applications

Conclusion At the 1989 annual meeting of the U.S. Tissue Culture Associations, Ricahrd am, a leading investigator in the serum-free nutrient requirements of cultured cells, commented on the process of medium development. He noted that a survey of major media manufacturers revealed that, among the top selling mammalian cell culture media formulations, most were nearly thirty years old.This commentary is noteworthy considering the tremendous changes in cell culture understanding and derived applications which have emerged over these three decades. Fastidious cell types relatively unknown to investigators of the 1950s and 1960s are now being cultivated in defined, serum-free environments. Culture environments range from limiting dilution clonal recoveries to maintenance cultures approaching tissue densities. While research applications continue to predominate, applications of cell culture have expanded to the engineered production of biopharmaceuticals, to replacement of animal models for toxicology testing, and to the preservation, activation and expansion of human cells, tissues and organs.It is likely that future nutrient medium development will be predicated upon the design of a minimal number of defined formulations of relatively generic utility to a broad class of cell types. Analytical techniques derived from those described herein will be exploited in the user laboratory and in collaboration with the supplier to optimize the nutrient composition for the desired biological response.     

Efficient production of wheat-barley hybrids and preferential elimination of barley chromosomes

Summary Intergeneric hybridization between four common wheat cultivars, Triticum aestivum L. cultivars Chinese Spring, Norin 12, Norin 61, and Shinchunaga, and cultivated barley, Hordeum vulgare L. cultivars Betzes, Nyugoruden, Harunanijou, and Kinai 5 were carried out in a greenhouse under 15 – 20 °C and long-day (15 h) photoperiod conditions. Two days prior to pollination, a 100 mg/1 2,4-D solution was injected into wheat stems. Among wheat cultivars, Norin 12, Norin 61, and Shinchunaga showed higher crossabilities than that of Chinese Spring, suggesting the presence of crossability gene(s) other than the kr system of Chinese Spring. Variation was also found among the barley cultivars as male parents. Betzes barley showed the highest crossability with wheat. Thus, the cross Norin 12×Betzes showed the highest crossability (8.25%), followed by Norin 61 ×Betzes (6.04%), Shinchunaga×Betzes (5.00%), and Shinchunaga×Kinai 5 (5.00%). The embryos were rescued by culture at 15–20 days after pollination. Seventyfour plants were obtained from 82 embryos. The morphology of the hybrid plants resembled that of wheat parents. Among 60 seedlings observed, 28 had 28 chromosomes, 8 had 21, 23 had aneuploid numbers of chromosomes (22–27), and 1 had 29 chromosomes. About half of the aneuploid hybrids showed mosaicism for chromosome number. By analyzing five isozyme markers of barley chromosomes, the chromosome constitutions of the aneuploid hybrids were determined. Barley chromosomes 1 and 5 were found to be preferentially eliminated in the hybrids, while chromosomes 2 and 4 were eliminated infrequently. The conditions and genetic factors for high crossability and the tendency of barley chromosome elimination are discussed.     

矮牵牛组织培养及变异苗的RAPD分析

探索了矮牵牛组织培养的配方,对组培过程中产生的表型变异植株进行了DNA水平的分析。并对矮牵牛叶片组织培养及产生变异的影响因素等进行探讨。