银河I号杨叶外植体再生体系建立

以银河 号杨 (Populus alba× P.hopeiensis)无菌试管苗叶片为外植体 ,建立了叶片外植体快速繁殖的培养程序。实验结果表明 ,MS附加 6 - BA∶ IAA(5∶ 1 )、每天 1 4h光照 ,可诱导离体叶片产生大量不定芽 ,且生根试管苗叶片的不定芽分化率明显高于无根试管苗。不定芽长至 1 cm以上时切下后转至 1 /2 MS附加 IBA 0 .2 mg/L的培养基上 ,可生根而形成完整的再生植株     

陆地棉子叶节高效再生体系的建立

对3个陆地棉材料子叶节进行离体培养,建立了高效的再生体系.其中,启动培养基为MSB+2.5 mg/L 6-BA +2.5 mg/LKT,芽分化培养基为MSB+0.3 mg/L 6-BA +0.3 mg/LKT,通过器官发生途径诱导形成丛生芽,芽伸长培养基MSB.以上培养基均含葡萄糖糖30 mg/L.此再生体系每个外植体可获得2.75个以上不定芽,嫁接移栽驯化后,成活率接近95%.     

茎杆的第一节间离体再生空心菜

以空心菜（Ipomoea aquatica Forsk.）品种‘白骨柳叶’为材料,通过筛选植物外植体和调整培养基激素配比等方法,首次建立了茎秆第一节间为外植体的空心菜离体再生体系。结果表明,在MS基本培养上添加0.05% 的植物组织培养抗菌剂PPM可获得大量空心菜无菌苗;植株子叶和下胚轴的切段均未诱导出不定芽,而茎秆第一节间为外植体能成功诱导出不定芽,诱导成功率为20%,最佳培养基配方为MS+1.0 mg/L 6-BA+0.1 mg/L IAA;不定芽诱导生根的最佳培养基配方为1/2 MS+0.1 mg/L NAA,生根率为100%。诱导成功后,将完整的再生苗移栽至基质土中,成活率可达100%。     

黄瓜子叶节高频再生体系建立及再生植株倍性观察

以‘农城3号’黄瓜子叶节为试材,研究不同苗龄、不同激素对黄瓜离体植株再生频率及再生不定芽的影响,并对再生植株进行了根尖染色体计数和叶片气孔保卫细胞叶绿体数目的鉴定。结果表明,(1)6-BA对黄瓜不定芽诱导起关键作用,IBA不利于黄瓜不定芽的诱导。(2)MS 2.0mg/L6-BA培养基是‘农城3号’黄瓜通过子叶节进行不定芽诱导再生植株的最佳培养基。(3)正常发育的无菌苗4~5d左右子叶不定芽再生频率较高,苗龄超过5d,不定芽再生频率明显下降。黄瓜子叶再生植株与实生苗的根尖染色体数目均为2n=14,再生植株与实生苗的气孔保卫细胞叶绿体数均分布在6~10的范围内,说明再生植株的倍性没有发生改变。     

银河Ⅰ号杨叶外植体再生体系建立

以银河Ⅰ号杨(Populus alba × P.hopeiensis)无菌试管苗叶片为外植体,建立了叶片外植体快速繁殖的培养程序.实验结果表明,MS附加6-BA∶IAA(5∶1 )、每天14 h光照,可诱导离体叶片产生大量不定芽,且生根试管苗叶片的不定芽分化率明显高于无根试管苗.不定芽长至1 cm以上时切下后转至1/2 MS附加IBA 0.2 mg/L的培养基上 ,可生根而形成完整的再生植株.     

大豆再生体系的研究

选用东北地区主栽品种‘黑农41’的子叶节和胚尖作为外植体,建立了‘黑农41’的子叶节和胚尖再生体系,并对2种再生体系进行了比较.结果表明子叶节最适芽诱导培养基为MSB5+1.0 mg/L6-BA+0.2 mg/LIBA,最适生根培养基为MSB5+1.0mg/LIBA;胚尖最适芽诱导培养基为MSB5 +0.2 mg/L6-BA +0.2 mg/LIBA,最适生根培养基为MSB5+2.0 mg/LIBA,子叶节更适合作为遗传转化的受体材料.     

西兰花再生体系的建立与优化

目的:以西兰花无菌苗为材料,对影响西兰花再生的各种素进行研究,建立并优化西兰花的再生体系.方法:用组织培养法,对西兰花的外植体进行诱导生芽、生根.结果:不同品种的再生率差别较大,珠绿和青秀两个品种的再生率达91%以上,而鼎丰一号仅为80.3%;培养6 d 的下胚轴分化能力最强;当培养基中添加3 mg/L 6-BA 和0.2 mg/L IAA 时,下胚轴的分化率最高,玻璃化程度也降到最低;当培养基中添加0.2 mg/L IBA 或0.2 mg/L NAA时,可成功诱导不定芽生根.结论:建立并优化了西兰花再生体系,为西兰花遗传转化体系的建立奠定了基础.     

猕猴桃实生苗再生体系的建立

以成熟饱满的美味猕猴桃种子发芽获得实生苗,分别以实生苗的茎段、叶柄和叶片为材料建立了再生体系。结果表明:种子以2.5mg/L的赤霉素(GA3)处理8h较适合;以培养基发芽较为适合;茎段、叶柄和叶片的愈伤组织的诱导率均为100%,且茎段、叶柄比叶片容易脱分化,但叶片的平均出苗率最高。再生苗在移栽5d后,开始长出新叶,10d后就能完全适应外界环境。     

毛华菊3种瓣型株系再生体系的建立

毛华菊(Chrysanthemum vestitum)是栽培菊花(C.×morifolium)的近缘六倍体野生种之一,其自然群体中的舌状花与栽培菊花一样具有典型的平瓣型、管瓣型和混合瓣型变异,是研究菊属植物瓣型变异的理想材料。其舌状花发育过程受生长素和花器官发育关键差异基因的影响,但目前缺乏稳定高效的不同瓣型毛华菊株系的再生体系,制约了毛华菊瓣型相关基因的研究。利用在河南省伏牛山收集的3种瓣型毛华菊株系,以叶片和茎间薄层为外植体建立再生体系。结果表明,以平瓣型叶片为外植体,其愈伤组织诱导和不定芽分化最佳培养基为MS+1 mg·L^-1 NAA+2mg·L^-1 6-BA,接种20天愈伤组织诱导率为100%,不定芽分化率达100%;最佳生根培养基为1/2MS+0.2 mg·L^-1 NAA,生根率达100%。平瓣型毛华菊的叶片最佳再生体系也适用于管瓣型和混合瓣型株系,不定芽分化率分别为83.46%和91.67%,生根率均为100%。移栽后对开花植株进行观察,发现利用叶片再生体系获得的3种不同瓣型再生植株花型稳定,为后续利用不同瓣...     

甜高粱再生体系的建立

利用甜高粱成熟种子和成熟胚作为外植体, 通过愈伤组织再生途径建立了植株再生体系。结果表明, 成熟种子在添加1.38 g.L-1脯氨酸、500 mg.L-1水解酪蛋白和3.0 mg.L-1 2, 4-D的MS培养基上可以较快地诱导出生长状态良好的愈伤组织;成熟胚在添加1.38 g.L-1脯氨酸、500 mg.L-1水解酪蛋白、2.5 mg.L-1 2, 4-D和0.1 mg.L-1KT的MS培养基上也能诱导出生长良好的愈伤组织。将愈伤组织转移到MS+1 mg.L-1 IAA + 0.5 mg.L-1 6-BA培养基上诱导芽, 然后再转移到MS+3 mg.L- 1IBA培养基上生根后, 可发育成为完整的植株。     

Eleven new microsatellites for hop (Humulus lupulus L.)

We present a new set of 11 polymorphic microsatellite primer sequences for use with Humulus lupulus. Microsatellite‐enriched libraries for GA_n and GT_n types of repeats were produced. Sequencing of 72 clones revealed 42 unique inserts containing microsatellites, out of which 19 primer pairs were designed and microsatellite amplification was tested on 39 wild hops and cultivars. Eleven primer pairs showed single locus amplification with 2–13 alleles, average 7.2, of which 17 unique alleles were discovered. One primer pair amplified too strong stutter bands, one locus was monomorphic and multilocus amplification was obtained with the remaining six primer pairs.     

啤酒花茎段培养及快速繁殖技术研究

以啤酒花的幼嫩茎段作为外植体,研究出理想的一次性成苗培养基：ＭＳ＋６－ＢＡ０．０１ｍｇ／Ｌ＋ＩＡＡ０．１ｍｇ／Ｌ＋琼脂６．５ｇ／Ｌ＋葡萄糖２％。试管苗月增殖率４．２,生根率１００％,成苗率９０％。采用试管苗微扦插的方式进行快速繁殖,经锻炼后直接移入经改造后的土壤中,移栽成活率达９３．８％。     

Cryopreservation of in vitro-grown shoot-tips of hop (Humulus lupulus L.) using encapsulation/dehydration

 A cryopreservation procedure using encapsulation/dehydration was established for shoot-tips obtained from in vitro-grown shoots of hop. After dissection, shoot-tips were encapsulated in medium with alginate and 0.5 M sucrose. Optimal conditions consisted of preculture for 2 days in solid medium with 0.75 M sucrose, or in increasing sucrose concentrations, desiccation for 4 h with silicagel in a flow cabinet (16% water content) followed by rapid freezing and slow thawing. Shoot recovery after freezing 60 min in liquid nitrogen was around 80%. No phenotypical changes were observed in the recovered plants from cryopreserved shoot-tips growing in the field. Received: 20 April 1997 / Revised: 20 February 1998 / Accepted: 1 Dezember 1998     

Isolation and characterization of polymorphic microsatellites for assessment of genetic variation of hops (Humulus lupulus L.)

Hop (Humulus lupulus L.) cultivars are vegetatively propagated and it is difficult to differentiate them during the process of propagation. Fingerprinting with molecular markers based on DNA could be a useful means of identifying different cultivars. Simple sequence repeats, or microsatellite markers, are the most suitable marker for genetic fingerprinting because they are multi‐allelic and co‐dominant. For this purpose, we have developed primers for 10 new polymorphic microsatellite loci that are suitable for genetic fingerprinting in hop.     

Enzymatic Reactions by Five Chalcone Synthase Homologs from Hop (Humulus lupulus L.)

The enzyme activities encoded in five cDNAs for chalcone synthase (CHS) homologs from hop were investigated. Only valerophenone synthase (VPS) and CHS_H1 showed both naringenin-chalcone and phlorisovalerophenone forming activity. Narigenin-chalcone production by VPS was much lower than by CHS_H1. Therefore, it is highly possible that flavonoid depends mainly on CHS_H1, while bitter acid biosynthesis depends mainly on VPS and CHS_H1.     

New polymorphic dinucleotide and trinucleotide microsatellite loci for hop Humulus lupulus L

One hundred and thirty-five microsatellite markers were developed for hop Humulus lupulus L. from di- and trinucleotide-enriched libraries. Seventy-eight primers showed amplification in two tested genotypes. Twenty-four loci were further characterized on a population of 34 hop samples and the number of alleles per locus, observed heterozygosity and expected heterozygosity ranged from two to 20 (9.7 on average), from 0.0294 to 0.9412 (0.6234 on average) and from 0.0294 to 0.9170 (0.6720 on average), respectively. These microsatellite markers will be further used for studying population structures and relationships and for identifying important qualitative and quantitative loci of hop.     

Modulation of breast cancer cell survival by aromatase inhibiting hop (Humulus lupulus L.) flavonoids

Hop flavonoids are being regarded as attractive molecules to prevent or treat certain forms of cancer. Studies have focused mainly on xanthohumol, the most abundant prenylated chalcone existing in hops extract. However, during the production of beer, or after its ingestion, xanthohumol originates different metabolites, among which isoxanthohumol and 8-prenylnaringenin. The aim of this work was to study the effect of the prenylflavonoids xanthohumol, isoxanthohumol and 8-prenylnaringenin on the breast cancer Sk-Br-3 cell line proliferation, apoptosis and activity of the enzyme aromatase (estrogen synthase). Aromatase activity was determined by a tritiated water assay, cell proliferation was assessed by [³H]thymidine incorporation, sulforhodamine B protein measurement and Ki-67 immunostaining and apoptosis was determined by TUNEL. Our results show that all tested prenylflavonoids were able to inhibit aromatase activity and thus, estrogen formation. Additionally, breast cancer cell line proliferation was decreased and apoptosis induced by all three compounds. The presence of 17β-estradiol in treatment medium was able to revert the effect of the prenylflavonoids on cellular proliferation. These observations strengthen the idea that hop flavonoids may have anti-breast cancer effects and shed new light on a possible mechanism of action by which these effects occur, namely through their ability to decrease estrogen synthesis.     

多效唑（MET）对啤酒花试管苗生长和移栽的影响

在培养基中加入0.1-0.2mg/L浓度的多效唑（MET）,啤酒花试管苗植株矮壮,单株生根数量增加、移栽成活率提高13％。0．5－2．0mg/L时,试管苗基部愈伤组织增多,枝叶失绿,生长受到抑制。初步发现多将唑对啤酒花试管苗的效力与葡萄糖用量呈正相关。     

Adventitious shoot regeneration in cultures of Humulus lupulus L. (hop) cvs. Brewers Gold and Nugget

 A very efficient protocol for plant regeneration from two commercial Humulus lupulus L. (hop) cultivars, Brewers Gold and Nugget has been established, and the morphogenetic potential of explants cultured on Adams modified medium supplemented with several concentrations of cytokinins and auxins studied. Zeatin at 4.56 μm produced direct caulogenesis and caulogenic calli in both cultivars. Subculture of these calli on Adams modified medium supplemented with benzylaminopurine (4.4 μm) and indolebutyric acid (0.49 μm) promoted shoot regeneration which gradually increased up to the third subculture. Regeneration rates of 60 and 29% were achieved for Nugget and Brewers Gold, respectively. By selection of callus lines, it has been possible to maintain caulogenic potential for 14 months. Regenerated plants were successfully transferred to field conditions. Received: 10 March 1997 / Revision received: 12 November 1997 / Accepted: 22 November 1998     

Organogenesis from internode-derived nodules of Humulus lupulus var. Nugget (Cannabinaceae): histological studies and changes in the starch content

The sequence of histological and histochemical events occurring during organogenesis from Humulus lupulus var. Nugget internode-derived nodules was studied. Sections were made and studies were carried out from the start of culture treatment until the development of shoot buds. Cell division was observed in both cambial and cortical regions during the first week of culture establishment. Cell division in cortical cells led to the formation of an incipient callus tissue. From the calluses prenodular structures of cambial origin appeared and gave rise to nodules from which shoot buds formed. Nodules kept separating into "daughter nodules" from which arose an increasing number of shoot buds. Iodide staining showed a strong starch accumulation in callus tissue and in prenodular structures. During shoot-bud primordia formation starch content decreased in nodules. Some starch was also noted in control explants (cultured on basal medium), however at a lower level than that observed in explants cultured on media with growth regulators. Shoot-bud regeneration was not observed in control explants.