水稻植株再生与农杆菌介导的遗传转化研究

本研究以不同籼稻品种的成熟胚为材料，研究了不同培养基对愈伤组织诱导、分化和植株再生的影响，并研究了利用农杆菌介导bar基因和ipt基因的遗传转化技术。结果表明，采用NBD(N6salts B5Vitamin casamino acids 500mg／L proline 500mg／L glutamine 300mg／L mannitol 36．4g／L sucrose 30g／L 2，4-D 2mg／L 6-BA 0．2mg／L phytagel 2．5g／L，pH 5．8)为诱导培养基，MSD(MS casamino acids 500mg／L proline 500mg／L glutamine 300mg／L mannitol 36．4g／L sucrose 30g/L 2，4-D 2mg／L 6-BA 0．2mg／L phytagel 2．5g／L，pH 5．8)与NBD培养基交替继代培养，籼稻愈伤组织诱导率高、质量较好；采用MSR3^e(MS casamino acids 500mg／L glutamine 300mg／L proline 500mg／L mannitol 36．4g／L maltose 30g／L TDZ 0．2mg／L phytagel 5g／L pH 5．8)为分化培养基，愈伤组织分化率均在90％以上。本研究利用农杆菌介导，将Act启动子驱动的抗除草剂基因(bar)和异戊烯基转移酶基因(ipt)构建的融合基因表达载体对水稻进行遗传转化。初步确定了愈伤组织筛选及分化培养基附加除草剂(glufosinate ammoniun)4mg／L、头孢霉素300mg／L，愈伤组织预培养2-3天，共培养基中加入乙酰丁香酮浓度为39mg／L，共培养2-3天。通过上述培养条件，已获得移栽成活的水稻转基因植株。     

小麦转基因再生植株培养体系的优化

以5个基因型不同的小麦幼胚，幼穗作为供试材料，探讨了品种基因型、ABA咱生根条件等对小麦离体培养的影响。结果表明，小麦品种豫麦18－64为遗传转化的良好受体材料；0．2mg/L NAA和0．8mg/L IAA可促进幼苗生根；在幼胚愈伤组织诱导前14d，用0．5mg/L ABA处理可有效抑制出芽率，并有利于促进胚愈伤组织形成和再生能力的保持.     

基因枪轰击后大麦幼胚的组织培养及植株再生研究

以基因枪轰击后的6个大麦品种的幼胚为材料，研究了不同质量浓度的ABA，2，4-D，ZT和IAA对胚幼的出愈率、愈伤组织分化特性和植株再生能力的影响。结果表明：用1．0mg／L的ABA在诱导培养基上处理幼胚能大大降低胚芽鞘发生率，并对出愈率和胚性愈伤组织的形成无不良影响；经1．0mg／L的2，4-D诱导产生的胚性愈伤组织转到分化培养基上以后，芽分化率可达8％～17％，而4．0mg／L 2，4-D处理芽分化率极低。1．0mg/L ZT与0．1mg／L IAA配比能降低分化过程中不定根的发生率，有利于芽的分化。通过植株再生系统的优化，6个大麦品种的转化幼胚植株再生率达到2％～8％。     

梅花品种"美人"叶片离体再生体系建立

以“美人’梅离体培养的叶片为材料进行愈伤组织诱导再生研究，诱导过程分两步，第一步是愈伤组织的诱导，诱导过程中生长素2,4-D起了决定性的作用，并且以液体培养基为佳，而且暗培养的愈伤组织再分化率明显高于光培养。具体培养基为1／2WPM＋5mg／L 2,4-D；第二步是在愈伤组织诱导出之后，进一步诱导愈伤组织再生出植物，在经过7d液体培养基的诱导之后将离体叶片转入1／2WPM＋1．0mg／L 6-BA＋0．1mg／L NAA＋8mg／L AgNO3＋100mg／L水解乳蛋白培养，再分化率可以达到24．25％。愈伤组织分化出苗之后，进一步诱导生根，“美人”梅最佳生根培养基是1／2WPM＋0．5mg／L NAA。     

培养基和培养时间对水稻成熟和胚培养力的影响

本试验研究了10种不同成份的培养基以及延长诱导培养时间对水稻成熟种胚培养力的影响。结果表明,水稻品种对不同培养基的反应是不同的。以∠S—5(∠S基本培养基+1.5mg/L2.4—D+1mg/1 KT+2%蔗糖+0.8%琼脂,pH5.8)培养基的愈伤组织诱导率为最高,诱导的愈伤组织质量较好。诱导的愈伤组织其绿苗分化率也较高,诱导培养60天其愈伤组织仍具有较高的分化能力。     

杂花苜蓿高效再生体系的建立

通过对新疆塔城杂花苜蓿品种下胚轴再生植株的系统研究，经筛选获得种子萌发培养基为：1／2MS无糖培养基；胚性愈伤组织诱导培养基：MS＋2，4-D2mg／L＋6-BA0．5mg／L＋蔗糖3％。胚性愈伤组织诱导率为100％；胚状体诱导培养基：SH＋2，4-DZing／L＋6-BA0．5mg／L＋CH250mg/L＋蔗糖1％，胚状体诱导率为86．96％；胚状体萌发培养基：SH＋GA30．5mg／L＋CH250mg／L＋蔗糖1％，胚状体萌发率为90％-95％；生根培养基：1／2MS＋IBA0．5rag／L＋蔗糖1％。生根率为100％。完成再生时间120d左右。     

转玉米C4型pepc水稻成熟胚为外植体的水稻悬浮细胞系的优化

为了研究外源玉米光合作用关键酶C4型磷酸烯醇式丙酮酸羧化酶(phosphoenolpyruvate carboxylase,PEPC)基因在C3植物水稻中提高光合效率的分子机理,本研究以高表达玉米C4型pepc水稻(PC)和未转基因野生型Kitaake (WT)为材料,探索高效且稳定水稻悬浮细胞系建立的方法.结果表明:PC成熟胚愈伤组织的诱导培养基中添加2 mg/L2,4-D和1 mg/L 6-BA,可出诱导出高频的愈伤组织；再通过3次继代培养(2～0.5 mg/L 2,4-D,2～0.2 mg/L 6-BA,1 mg/L ABA),逐步降低2,4-D和6-BA的浓度,可获得了疏松、颗粒状且分散的愈伤组织块；然后转接到水稻悬浮细胞液体培养基中(0.5 mg/L2,4-D,0.2 mg/L 6-BA和1 mg/LABA),在28℃下培养,新原液的比例为3∶1,经42 d,可建立稳定且高效的供试水稻悬浮细胞系.     

欧洲七叶树体细胞胚胎发生研究

以欧洲七叶树幼嫩叶片为外植体，在离体条件下成功诱导出体细胞胚胎(体胚)。研究结果表明，诱导愈伤组织的适宜培养基是MS 2，4-D 2mg／L KT0．2mg／L，MS BA 8mg／L NAA 1mg／L有利于胚性愈伤的诱导和增殖，添加ZT2mg／L或BA 5mg／L和NAA0．2mg／L的MS培养基有利于体胚发育和成熟。     

粳稻恢复系C418成熟胚转基因再生体系的建立

为了建立粳稻恢复系C418转基因再生体系,本研究以不同培养基、不同2,4-D浓度、不同KT浓度及不同6-BA浓度的不同组合培养基来诱导C418成熟胚愈伤组织,以获得最适C418诱导的培养基,然后以获得的愈伤组织利用农杆菌介导法进行转基因再生体系的建立。结果表明:NBD培养基为6种培养基中最适培养基;2,4-D浓度为2.0 mg/L时,诱导率最高;随着6-BA的浓度增加诱导率降低;KT对C418成熟胚愈伤组织的诱导没有影响。本研究成功建立了C418转基因再生体系。     

水稻植株再生与农杆菌介导的遗传转化研究

本研究以不同籼稻品种的成熟胚为材料, 研究了不同培养基对愈伤组织诱导、分化和植株再生的影响,并研究了利用农杆菌介导bar基因和ipt基因的遗传转化技术.结果表明,采用NBD (N6salts B5Vitamin casamino acids 500 mg/L proline 500 mg/L glutamine 300 mg/L mannitol 36.4 g/L sucrose 30 g/L 2,4-D 2 mg/L 6-BA 0.2 mg/L phytagel 2.5 g/L,pH 5.8) 为诱导培养基,MSD(MS casamino acids 500 mg/L proline 500 mg/L glutamine 300 mg/L mannitol 36.4 g/L sucrose 30 g/L 2,4-D 2 mg/L 6-BA 0.2 mg/L phytagel 2.5 g/L,pH 5.8)与NBD培养基交替继代培养,籼稻愈伤组织诱导率高、质量较好;采用MSR3c(MS casamino acids 500 mg/L glutamine 300 mg/L proline 500 mg/L mannitol 36.4 g/L maltose 30 g/L TDZ 0.2 mg/L phytagel 5 g/L pH 5.8)为分化培养基,愈伤组织分化率均在90%以上.本研究利用农杆菌介导,将Act启动子驱动的抗除草剂基因(bar)和异戊烯基转移酶基因(ipt)构建的融合基因表达载体对水稻进行遗传转化.初步确定了愈伤组织筛选及分化培养基附加除草剂(glufosinate ammoniun)4 mg/L、头孢霉素300 mg/L,愈伤组织预培养2-3天,共培养基中加入乙酰丁香酮浓度为39 mg/L,共培养2-3天.通过上述培养条件,已获得移栽成活的水稻转基因植株.     

Location of a high-lysine gene and the DDT-resistance gene on barley chromosome 7

Summary The high-lysine gene in Risø mutant 1508 conditions an increased lysine content in the endosperm via a changed protein composition, a decreased seed size, and several other characters of the seed. The designation lys3a, lys3b, and lys3c, is proposed for the allelic high-lysine genes in three Risø mutants, nos 1508, 18, and 19. Linkage studies with translocations locate the lys3 locus in the centromere region of chromosome 7. A linkage study involving the loci lys3 and ddt (resistance to DDT) together with the marker loci fs (fragile stem), s (short rachilla hairs), and r (smooth awn) show that the order of the five loci on chromosome 7 from the long to the short chromosome arm is r, s, fs, lys3, ddt. The distance from locus r to locus ddt is about 100 centimorgans.     

A note on the origin of Kalanchoë x vadensis Boom & Zeilinga (Crassulaceae)

Summary K x vadensis is a hybrid of K. blossfeldiana and K. marmorata obtained after doubling the number of chromosomes.     

Simultaneous Determination of Four Phenolic Acids in Radix Salviae Miltiorrhizae Formula Granules by QAMS

[Objectives]This study aimed to establish a QAMS(quantitative analysis of multi-components by single-marker)method for simultaneous determination of four phenol...     

Avoidance of leaf rust fungi in wild relatives of cultivated cereals

Summary Avoidance of rust fungi that was based on poor appressorium induction was previously found in Hordeum chilense. In the present study 95 accessions of Triticeae were screened for avoidance of Puccinia hordei. The percentage of appressorium formation per germinated spore ranged from 6 to 90%. On none of the 41 accessions of Aegilops, Agropyron, Elymus, Secale, Thinopyrum or Triticum studied was the rate of appressorium formation lower than 25%. Lower rates of appressorium formation were, however, found on accessions of wild barley species Hordeum brachyantherum, H. marinum, H. parodii and H. secalinum. Its implications in cereal breeding are discussed.     

Present status and future strategy in breeding faba beans (Vicia Faba L.) for resistance to biotic and abiotic stresses

Progress is being made, mainly by ICARDA but also elsewhere, in breeding for resistance to Botrytis, AScochyta, Uromyces, and Orobanche; and some lines have resistance to more than one pathogen. The strategy is to extend multiple resistance but also to seek new and durable forms of resistance. Internationally coordinated programs are needed to maintain the momentum of this work.Tolerance of abiotic stresses leads to types suited to dry or cold environments rather than broad adaptability, but in this cross-pollinated species, the more hybrid vigor expressed by a cultivar, the more it is likely to tolerate various stresses.     

Water Extraction Process of Chinese Herbal Compound Man Gan Ning

[Objectives]To optimize the water extraction process of Chinese Herbal Compound Man Gan Ning and establish a method for its extraction and content determination...     

Cytoplasmic male sterility,resistance to gall mite and mildew,and season of leafing out in black currants

Summary Cytoplasmic male sterility (cms) is described in the F₁ hybrids Ribes × carrierei (R. glutinosum albidum × R. nigrum) and R. sanguineum × R. nigrum. In backcrosses to R. nigrum, progenies with R. glutinosum cytoplasm were either all male sterile, or segregated for full male fertility (F) and complete (S) and partial (I) male sterility. Ratios of F:I+S suggested that two linked genes controlled cms, F plants being dominant for one (Rf ₁) and recessive for the other (Rf ₂).Segregation for cms in relation to three linded genes, Ce (resistance to the gall mite, Cecidophyopsis ribes), Sph ₃(resistance to American gooseberry mildew, Sphaerotheca mors-uvae) and Lf ₁(one of two dominant additive genes controlling early season leafing out) indicated that Rf ₁and Rf ₂were in this linkage group. The gene order and approximate crossover values appeared to be: % MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXafv3ySLgzGmvETj2BSbqef0uAJj3BZ9Mz0bYu% H52CGmvzYLMzaerbd9wDYLwzYbItLDharqqr1ngBPrgifHhDYfgasa% acOqpw0xe9v8qqaqFD0xXdHaVhbbf9v8qqaqFr0xc9pk0xbba9q8Wq% Ffea0-yr0RYxir-Jbba9q8aq0-yq-He9q8qqQ8frFve9Fve9Ff0dme% aabaqaciGacaGaamqadaabaeaafaaakeaacaWGdbGaamyzamaamaaa% baGaaiiiaiaacccacaGGWaGaaiOlaiaacgdacaGG0aGaaiiiaiaacc% caaaGaaiiiaiaacccacaGGGaGaamOuaiaadAgaliaaigdakmaamaaa% baGaaiiiaiaacccacaGGGaGaaiiiaiaaccdacaGGUaGaaiOmaiaacs% dacaGGGaGaaiiiaiaacccacaGGGaGaaiiiaaaacaWGsbGaamOzaSGa% aGOmaOWaaWaaaeaacaGGGaGaaiiiaiaacccacaGGGaGaaiiiaiaacc% cacaGGGaGaaiiiaiaacccaaaGaamitaiaadAgaliaaigdakmaamaaa% baGaaiiiaiaacccacaGGGaGaaiiiaiaacccacaGGGaGaaiiiaiaacc% cacaGGGaGaaiiiaiaacccacaGGGaaaaiaadofacaWGWbGaamiAaSGa% aG4maaaa!6E4D!\[Ce\underline { 0.14 } Rf1\underline { 0.24 } Rf2\underline { } Lf1\underline { } Sph3\]. Crossover values of 0.36 for Ce-Lf ₁, and 0.15 for Lf ₁-Sph ₃were estimated from the relative mean differences in season of leafing out between seedlings dominant and recessive for Ce and Sph ₃.It is suggested that competitive disadvantage of lf ₁-carrying gametes and/or zygotes at low temperatures may be implicated in the almost invariable deficit of plants dominant for the closely linked mildew resistance allele Sph ₃. Poor performance of lf ₁- (and possibly lf ₂-) carrying gametes and young zygotes during periods of low temperature at flowering might also account for the liability of some late season cultivars and selections to premature fruit drop (running off).     

Optimization of Extraction Technology and Determination of Content of Total Phenols of Leaves in Acanthopanax giraldii Harms

[Objectives] To determine the optimum extraction technology for total phenols of leaves in Acanthopanax giraldii Harms.[Methods]The single factor test and ortho...     

Pollen and pollination experiments. III. The viability of apple and pear pollen as affected by irradiation and storage

Summary Apple and pear pollen was irradiated with doses of 0, 50, 100, 250 and 500 krad (gamma rays) and stored at 4°C and 0–10% r.h. From the in-vitro germination percentages an average LD 50 dose of about 220 krad was estimated. For both irradiated and untreated pollen a close and corresponding lineair relationship existed between germination percentage and pollen tube growth.Irradiated pollen was much more sensitive to dry storage conditions than untreated pollen, resulting in less germination and more bursting. Apparently, irradiation caused the pollen cell membrane to lose its flexibility faster than normal. Rehydration of dry-stored, irradiated pollen in water-saturated air restored germination percentages up to their initial levels. The importance of this procedure in germination trials is stressed.     

Screening techniques and sources of resistance to parasitic angiosperms

Parasitic angiosperms cause great losses in many important crops under different climatic conditions and soil types. The most widespread and important parasitic angiosperms belong to the genera Orobanche, Striga, and Cuscuta. The most important economical hosts belong to the Poaceae, Asteraceae, Solanaceae, Cucurbitaceae, and Fabaceae. Although some resistant cultivars have been identified in several crops, great gaps exist in our knowledge of the parasites and the genetic basis of the resistance, as well as the availability of in vitro screening techniques. Screening techniques are based on reactions of the host root or foliage. In vitro or greenhouse screening methods based on the reaction of root and/or foliar tissues are usually superior to field screenings and can be used with many species. To utilize them in plant breeding, it is necessary to demonstrate a strong correlation between in vitro and field data. The correlation should be calculated for every environment in which selection is practiced. Using biochemical analysis as a screening technique has had limited success. The reason seems to be the complex host-parasite interactions which lead to germination, rhizotropism, infection, and growth of the parasite. Germination results from chemicals produced by the host. Resistance is only available in a small group of crops. Resistance has been found in cultivated, primitive and wild forms, depending on the specific host-parasite system. An additional problem is the existence of pathotypes in the parasites. Inheritance of host resistance is usually polygenic and its transfer is slow and tedious. Molecular techniques have yet to be used to locate resistance to parasitic angiosperms. While intensifying the search for genes that control resistance to specific parasitic angiosperms, the best strategy to screen for resistance is to improve the already existing in vitro or greenhouse screening techniques.