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.     

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.     

Inactivation and elimination of human enteric viruses by Pacific oysters

Aims: To investigate the comparative elimination of three different human enterically transmitted viruses [i.e. hepatitis A virus (HAV), norovirus (NoV) and poliovirus (PV)] and inactivation of HAV and PV by Pacific oysters. Methods and Results: New Zealand grown Pacific oysters (Crassostrea gigas) were allowed to bioaccumulate HAV, NoV and PV. Samples of oyster gut, faeces and pseudofaeces were then analysed by using real‐time RT‐PCR to determine the amount of viral RNA and cell culture methods to identify changes in the number of plaque forming units. The results suggest that the majority of the PV present in the oyster gut and oyster faeces is noninfectious, while in contrast, most of the HAV detected in the oyster gut are infectious. Depuration experiments identified a large drop in the count of PV in the gut over a 23‐h cleansing period, whereas the levels of HAV and NoV did not significantly decrease. Conclusions: Human enterically transmitted viruses are eliminated and inactivated at different rates by Pacific oysters. Significance and Impact of Study: The research presented in this article has implications for risk management techniques that are used to improve the removal of infectious human enteric viruses from bivalve molluscs.     

苹果脱毒苗与常规苗脂肪酸组成与抗氧化酶活性的比较研究

以甘肃陇东黄土高原3个主栽苹果品种嘎拉、秦冠、富士的12代继代培养脱毒苗与常规苗进行叶片类脂脂肪酸组成及抗氧化酶活性的比较研究。结果表明,苹果脱毒的膜脂过氧化程度降低,叶片类脂脂肪酸不饱和度提高。过氧化氢酶(CAT)和过氧化物酶(POD)活性升高,而超氧化物歧化酶(SOD)活性与常规苗无明显差异。     

Patterns of fluctuating asymmetry in flowers: Implications for sexual selection in plants

Characters in animals used in signalling and subjected to strong directional selection often demonstrate (i) an elevated level of fluctuating asymmetry (small random deviations from bilateral symmetry) and (ii) a negative relationship between the degree of individual fluctuating asymmetry and the size of a given character. We tested these two predictions in plants since flowers are subjected to strong directional selection and are involved in signalling to pollinators, whereas leaves are supposed not to be directly involved in signalling. The overall level of fluctuating asymmetry in a number of plant species with bilaterally or radially symmetric flowers was not generally higher in floral traits than in leaves. The level of fluctuating asymmetry in plants was sometimes significantly consistent within individuals. The absolute degree of individual fluctuating asymmetry in floral traits was generally negatively related to the size of the trait, while there was a positive relationship for leaves. The degree of individual fluctuating asymmetry in floral traits was marginally negatively related to the degree of individual fluctuating asymmetry in leaf traits. These patterns of fluctuating asymmetry in plants suggest that (i) the degree of asymmetry in flowers signals different aspects of quality than does the degree of asymmetry in leaves, and that (ii) fluctuating asymmetry in flowers often reflects the phenotypic quality of individual plants.     

Phytase production by high cell density culture of recombinant Bacillus subtilis

Bacillus subtilis BD170, harboring a plasmid pGT44[phyC] carrying the phytase gene (phyC) and a phosphate-depletion inducible pst-promoter, was grown in a 2 l bioreactor. Using a controlled feeding of glucose, high cell densities of 32 and 56 g dry cell weight l^–1 were achieved with peptone and yeast extract, respectively, as the complex nitrogen sources in a semi-defined growth medium. The fed-batch protocol was applied to production of recombinant phytase and a high extracellular phytase activity (48 U ml^–1) was reached with peptone. Although the yeast extract feeding resulted in a higher cell density, it was unsuitable as a medium component for phytase expression due to its relatively high phosphate content.     

The role of cholecystokinin (CCK8) on glucose production and elimination, and on plasma insulin and glucose in rats

The effects of the gastrointestinal hormone and neurotransmitter cholecystokinin (CCK8) are complex, since it exhibits both an insulinotropic and a glucagonotropic effect. We investigated CCK8 in vivo with respect to glucose fluxes (production and elimination) at both low (6 mM) and high plasma glucose levels (9 mM) using the primed constant -[3-³H]glucose infusion technique. In the presence of high glucose levels there was a dose-dependent increase in glucose elimination by CCK8. No effect of CCK8 on glucose production at a high glucose infusion rate (500 mg/kg per h) was observed in contrast to a low glucose infusion rate (100 mg/kg per h); plasma glucagon levels were elevated. All effects on glucose production and elimination were specific, since they were abolished by the CCK receptor antagonist L-364,718. In summary, glucose elimination was slightly increased by CCK8 at low glucose levels but increased to a greater extent at high glucose levels; glucose production was increased by CCK8 only at low glucose levels. Thus, CCK is a regulator of glucose homeostasis.