Introduction of rice chitinase gene into wheat via low energy Ar+ beam implantation

A chitinase gene (RCH8) in plasmid vector pCAMBIA1308 was delivered into 3 wheat cultivars (Yangmai 158, Wan 9210, Wanmai 32) by low energy Ar⁺ beam-mediated method. Preliminary calli from treated mature embryos were first selected on hygromycin (Hm, 20 or 30 mg/L) containing medium. After the resistant calli formed, they were transferred to the regeneration medium with 10 or 20 mg/L Hm. All the three wheat varieties obtained transgenic plants. PCR and PCR-Southern assays showed that most plants regenerated from the resistant calli were positive transgenic plants. Southern blot of the positive green plants confirmed stable integration of alien DNA into wheat genome. The plant transformation frequencies varied with the variety and ion dose implanted. Wanmai 32 possessed the highest transformation frequency, reaching 3.8% at a suitable implantation dose. The transformation frequency of Yangmai 158 and Wan 9210 varied from 0.5% to 2.5% and from 0.5% to 1.4%, respectively. Progeny test for resistance to wheat scab showed that the leaf extract of R₁ generation inhibited the growth of wheat scab strain R₀ and F₁₅.     

祁连龙胆愈伤组织和再生植株的生长及其两种药效成分分析

通过不同激素组合的培养基筛选出祁连龙胆（Gentiana Przewalskii Maxim.）的快速生长细胞系并获得再生植株. 采用反相高效液相色谱法测定愈伤组织及其再生植株中獐牙菜苦甙（Swertiamarin）、龙胆苦甙(Gentiopicroside)的含量. 结果表明：(1）祁连龙胆愈伤组织在不同培养基上诱导率和生长速度不同，在MS+2?mg/L 2,4 D +1?mg/L 6 BA +6?mg/L GA的培养基上诱导率最高，达78.0％. 在MS+3?mg/L 2,4 D +0.5?mg/L KT的培养基上生长最快，增殖倍数达到7.18； (2）祁连龙胆愈伤组织及其再生植株有效成分及含量与原植株相比均有差别，其中，愈伤组织含有0.265%的龙胆苦甙，低于原植株(4.225%)，再生植株含有0.279%的獐牙菜苦甙和1.579%的龙胆苦甙，前者高于原植株（0.260％），但后者略有下降. 按生长速度推算，愈伤组织及其再生植株积累的药效成分含量明显高于原植株，值得进一步开发利用.     

Hybrid plant regeneration from interfamilial somatic hybridization between grapevine (vitia vinifera L.) and Red Thorowax (Bupleurum scorzonerifolium willd)

The protoplasts of Red Thorowax ( Bupleurum scorzonerifolium) irradiated by ultraviolet light (UV) at an intensity of 260μW/cm2 for 0, 1,2 and 3 min respectively were fused with that of grapevine ( Vitia vinifera). The regenerated 19 clones, every one derived from a single fused cell, were identified as hybrids by phenotype, isozyme, chromosome and 5S rDNA spacer region analysis. The results reveal that all of them are somatic hybrids. 11 hybrid calli including asymmetric and symmetric products regenerated somatic embryos and young leaves after 5 months of culture, of which 4 hybrid cell lines derived from asymmetric fusion regenerated plants with roots after 8-10 months of culture. Inspection of chromosome showed that regeneration of whole plant was related to the decrease of chromosome number. Identification of 5S rDNA spacer region of the plants confirmed that they were interfamilial hybrid plants.     

抗除草剂旱稻转基因植株的获得

以旱稻丹粳旱５－５５、６－２４、６－３７的悬浮细胞及未成熟胚为受体材料，采用基因枪法将含ＢＡＲ基因的ｐＤＭ３０２质粒ＤＮＡ导入旱稻细胞中，经ＰＰＴ筛选获得抗性愈伤组织，筛选出的愈伤组织在分化培养基上再生出完整的旱稻转基因植株。Ｓｏｕｔｈｅｒｎ分子杂交分析表明，外源ＢＡＲ基因已整合到水稻基因组内，抗除草剂试验结果表明，转化植株对０．０１％Ｂａｓｔａ（有效成分为ＰＰＴ）有一定程度的抗性，说明外源ＢＡＲ     

Rice protoplasts isolated directly from mature-embryo-derived calli

Mature-embryo-derived calli of japonica rice (Oryza sativa L) Taipei 309 were used for replicated protoplast isolation experiments. Six out of nine callus lines produced protoplasts with satisfactory yield of 5.20×10⁶–8.96×10⁶ protoplasts/g FW (fresh weight). The remaining three callus lines initiated from seeds of cryopreserved-callus-derived plants had rooty calli, resulting in low yield of protoplasts and a large number of isolated banana-shape intact cells. Viability of protoplasts ranged 87.46%–94.15%. The average size of protoplasts was 207.49–379.04 μm² in different callus lines. Comparitive experiments were also carried out using both calli and suspension culture cells for protoplast isolation. The results demonstrated that protoplast isolation of calli was a substantially simplified and reliable method for preparing rice protoplasts. Jin Deming: born in Jan. 1959, Associated Professor     

农杆菌介导的DREB1A基因转化多花黑麦草及其转化体系的优化

采用携带卡那霉素抗性基因nptII和GUS基因的ubiquitin启动子驱动的表达载体pBI121／DREB1A的根癌农杆菌AGL1, 对多花黑麦草幼胚来源的胚性愈伤组织进行了遗传转化,并优化了各种影响因素。胚性愈伤组织经根癌农杆菌感染和共培养后,用50mg／L巴龙霉素筛选抗性愈伤组织,待抗性愈伤组织在IB分化培养基上分化成苗后用25mg／L卡那霉素进一步筛选再生植株, 获得了部分抗性植株。抗性植株的总DNA用DREB1A基因的特异引物进行PCR检测,转化频率为2.14％,PCR-Southern blot进一步验证了转化植株基因组中含有该外源基因。各种影响转化效率因素的优化实验表明,当转化时菌液浓度的OD600为2.0、侵染时间为1h、共培养时间为2d、共培养温度为21℃及在共培养期间使用乙酰丁香酮等,均可明显提高转化频率。     

应用农杆菌介导法的多年生黑麦草遗传转化研究

以草坪型多年生黑麦草Lolium perenne L.成熟种子为外植体,经过愈伤组织诱导和植株再生研究,建立了该草种的遗传转化体系,并成功地应用农杆菌介导法将克隆自辽宁碱蓬的甜菜碱醛脱氢酶基因(BADH)转移进多年生黑麦草,获得的转基因株系Gsc-LP5通过PCR检测证明了目的基因的存在,通过叶片离体检测法证明了作为检测基因随同质粒一同转入的抗潮霉素基因的表达,通过盆栽耐盐试验证明了转基因植株具有较强的耐盐能力.     

苏云金芽孢杆菌aiiA基因载体构建及石斛兰转化

 细菌性软腐病是石斛兰及其它兰花栽培和生产中的一大危害。苏云金芽孢杆菌Bacillus thuringiensis（Bt）中含有aiiA基因,该基因转入一些植物可赋予植物表现出软腐病抗性。从Bt菌中克隆出aiiA基因,并装载入植物表达载体pCAMBIA1301,构建出可用于转入石斛兰的pSAN载体;首次利用根癌农杆菌EHA105将pSAN载体转入石斛兰,并获得潮霉素抗性苗。这些抗性苗经GUS组织化学染色和PCR检测证实为转基因苗, 这将为获得具有软腐病抗性的石斛兰品种打下基础。     

Biogeochemical sequestration of carbon within phytoliths of wetland plants: A case study of Xixi wetland, China

As an important long-term terrestrial carbon sequestration mechanism, biogeochemical sequestration of carbon within phytoliths may play a significant role in the global carbon cycle and climate change. The aim of this study is to explore the potential of carbon bio-sequestration within phytoliths produced by wetland plants. The results show that the occluded carbon content of phytoliths in wetland plants ranges from 0.49% to 3.97%, with a CV (coefficient of variation) value of 810%. The data also indicate that the phytolith-occluded carbon (PhytOC) content of biomass for wetland plants depends not only on the phytolith content of biomass, but also the efficiency of carbon occlusion within phytoliths during plant growth in herb-dominated fens. The fluxes of carbon bio-sequestration within phytoliths of herb-dominated fen plants range from 0.003 to 0.077 t CO2 equivalents t-e-CO2ha-1a-1 . In China, 0.04×106 to 1.05×106t CO2 equivalents per year may be sequestrated in phytoliths of herbaceous-dominated fen plants. Globally, taking a fen area of 1.48×108 ha and the largest phytolith carbon biosequestration flux (0.077 t-e-CO 2 ha-1a-1 ) for herb-dominated fen plants, about 1.14×10 7 t CO 2 equivalents per year would have been sequestrated in phytoliths of fen plants. If other wetland plants have similar PhytOC production flux with herb-dominated fen plants (0.077 t-e-CO2ha-1a-1 ), about 4.39×10 7 t-e-CO2a-1 may be sequestrated in the phytoliths of world wetland plants. The data indicate that the management of wetland ecosystems (e.g. selection of plant species) to maximize the production of PhytOC have the potential to bio-sequestrate considerable quantities of atmospheric CO2 .     

毛花猕猴桃愈伤组织诱导与植株再生

从毛花猕猴桃（ＡｃｔｉｎｉｄｉａｅｒｉａｎｔｈａＢｅｎｔｈ）田间生长的茎段、种子无菌条件下萌发的下胚轴及试管苗的茎段和叶片得到愈伤组织。田间来源的茎段诱导愈伤组织较难，愈伤组织生长缓慢；下胚轴及试管苗茎段和叶片容易产生愈伤组织，愈伤组织生长旺盛。消毒时间、接种方法、基因型和培养基都可能影响到田间来源的茎段愈伤组织产生。在ＭＳ附加玉米素或６－苄氨基嘌呤（ＢＡＰ）的培养基上下胚轴来源的愈伤组织不经转代即分化芽和根。试管苗茎段和叶片在附加玉米素或Ｎ－（２－氯基－４－吡啶基）－Ｎ′－苯基脲（ＣＰＰＵ）的ＭＳ培养基上培养一代也分化出芽、试管苗茎段在附加０．００２５ｍｇ／ＬＣＰＰＵ和０．１ｍｇ／Ｌ吲跺乙酸（ＩＡＡ）的ＭＳ培养基上产生愈伤组织、芽的分化和苗生长都较理想；试管苗叶片则以附加０．０２５ｍｇ／ＬＣＰＰＵ和０．１ｍｇ／ＬＩＡＡ或０．５ｍｇ／Ｌ玉米素和０．１ｍｇ／ＬＩＡＡ的ＭＳ培养基较好。当苗伸长至１ｃｍ或以上时切下，转入ＭＳ基本培养基（大量元素减半）上后可形成完整植株。     

Inheritance of resistance to Helicoverpa armigera of 3 kinds of transgenic Bt strains available in upland cotton in China

There are 3 kinds of transgenic Bt strains, Shanxi 94-24, Zhongxin 94, and R19, in upland cotton in China. Their transgenic Bt insect-resistance cultivars or hybrids have been developed and grown by farmers. Genetic studies indicate that the resistance of the 3 transgenic Bt cotton strains to Helicoverpa armigera is controlled by one pair of non-allelic dominant genes. Linkage relationship between the resistant genes of R19 and Shanxi 94-24 transgenic Bt strains shows that they may be inserted in the same chromosome. F₁ hybrids crossed among the 3 strains show that high levels of protection from feeding damage are the same as that of their parents. Therefore, there is no co-suppression phenomenon in many transgenic plants. The results presented here afford a fundamental reliance in developing transgenic Bt insect-resistant cultivars and exploiting the heterosis of hybrids in upland cotton.     

Piezoelectric immunoassay of thyroidal triiodothyronine

A re-utilizable piezoelectric crystal (PC) immunosensor for thyroidal triiodothyronine (T3) has been developed. The crystal was coated with protein A and then reacted with T3 antibody. T3 could be detected in the range of 0.5×10⁻⁶ g/L to 10.0×10⁻⁶g/L by the, system. Crystal was regenerated by saturating with T3 and subsequent binding of a new anti-T3 antibody layer. The T3 assay could be repeated up to 5 times using each crystal. Supported by the National Natural Science Foundation of China Zha Hongying: born May, 1969, Ph. D.     

玉米与其近缘野生种杂交后代的组织培养

繁茂大刍草（Zea luxurians）和二倍体多年生类玉米（Zea diploperennis）是栽培玉米的野生近缘种，其抗逆性强，染色体数目也与玉米相等（2n=20），是玉米重要的种质资源。本工作通过远缘杂交、回交等技术将野生种染色体引入到玉米骨干自交系中，然后限幼胚诱导胚性愈伤组织并再生出大批植株；研究了基因型和培养基激素组合对胚性愈伤组织诱导、继代和植株再生的影响，以及体细胞再生植株的利用价值。     

Development of Triticum aestivum-Leymus racemosus ditelosomic substitution line 7Lr#1S（7A） with resistance to wheat scab and its meiotic behavior analysis

Leymus racemosus is highly resistant to wheat scab （Fusarum head bright）. The transfer of scab resistant gene from L. racemosus to Triticum aestivum is of great significance for broadening the base of wheat resistance. In the present study, the pollen of T. aestivum-L, racemosus monosomic addition line with scab resistance was treated by irradiation with 1200 R ^60Co-γ-rays prior to pollinating to emasculated wheat cv. Mianyang 85-45. Nine plants with a telocentric chromosome 7Lr#1S were observed in M1, and one ditelosomic substitution line 7Lr#1S was selected from selfcrossing progenies and confirmed by chromosome C-banding and GISH. Furthermore, a co-dominant EST-SSR marker CINAU 31 was employed to identify this substitution line. A pair of chromosome 7A of common wheat were found to be replaced by a pair of telocentric chromosome 7Lr#1S, and further investigation showed that chromosome configuration of the substitution line at MI of PMCs after GISH was 17.50Ⅱ^w ＋ 2.19Ⅱ^w ＋ 0.42Ⅱ^7Lr#1S ＋ 1.08 Ⅰ^7Lr#1S ＋ 0.69 Ⅰ^w. Two telocentric chromosomes paired as a bivalent in 59.7% of PMCs. Abnormal chromosome behaviors of telocentric chromosomes were observed in part of PMCs at anaphase I and telophase I, including the moving of two telocentric chromosomes to the same pole, lagging and earlier separation of their sister chromatid. All these abnormal behaviors can be grouped into three distinct types of tetrads according to different numbers of 7Lr#1S in their daughter cells and various micronucleus in some tetrads. However, due to the high transmission frequency of the female and male gametes with a 7Lr#1S, 84% of the selfcrossing progeny plants had ditelosomic substitution. The substitution line showed high resistance to wheat scab in a successive two-year test both in the greenhouse and field; hence, the line will be particularly valuable for alien gene mapping, small fragment translocation induction and telosomic cytological behavior analysis.     

利用基因枪法向高羊茅导入bar基因的研究

利用基因枪法,以bar基因转化高羊茅(Festuca arundinacea Schreb.)的下胚轴愈伤组织,对获得的再生植株进行的PCR检测和点杂交分析表明,外源bar基因片段已经整合至高羊茅基因组中.同时,对转基因植株进行的草丁膦涂抹实验发现,转基因植株对除草剂的耐性提高.     

Introducing antisense <Emphasis Type="Italic">waxy</Emphasis> gene into rice seeds reduces grain amylose contents using a safe transgenic technique

Rice (Oryza sativa L.) eating quality is one of themost important traits. Amylose content (AC) in rice en-dosperm is a major index affecting rice eating quality[1,2].It has a negative correlation with gel consistency of rice[3].Based on amylose content, r…     

Studies of Improving the Frequency of Indica Rice Transformation by Biolistic Bombardment

In order to improve the frequency of indica rice transformation by biolistic bombardment,suitable culture conditions for embryonic calli,an optimal selection scheme for resistant calli and seedling,and optimum bombardment parameters a investigated by using 14 commercially important indica rice cultivars.The main results show that the CC medium with 36g/L mannitol is a scheme subculture medium in which the browning of indica rice calli can be mitigated significantly;The concentration of 30-40mg/L Hyg or 150-200mg/L G418 or 10-20mg/L Basta is suitable for selection of resistant calli;The transformation parameters of 100μg gold powder absorbing 0.2μg DNA per shot and 900 psi helium pressure and 6 cm bombardment distance and bombarded twice for each plate give the best result;Keeping the target calli on osmotic medium containing 60g/L mannitol from 12-24h before bombardment to 24-48h after it can increase the efficiencies of transformation.Furthermore,some transgenic indica rice plants are obtained using this optimized transformation system.     

农杆菌介导的胆碱脱氢酶基因(betA)转化小麦及影响转化效率的因素

用LBA4404／pCDH，Agl 1／pUNN2和Ag;P(无质粒)3种菌株分别转化小麦品种济南177、99P、核生3号幼胚诱导的愈伤组织以及济南177的幼胚．其中以胚性愈伤组织为外植体，获得了转基因植株．PCR和PCR-Southem分析证实转化植株中包含了外源基因．通过染色体分析，发现胚性愈伤组织在农杆菌LBA4404侵染后形成的染色体削减比经Agl1侵染和未经感染的对照形成的染色体削减程度更大，甚至发现较多的染色体断片．分析了农杆菌菌株，材料的基因型，外植体类型，培养时间和温度以及筛选的时间和周期对于转化效率的影响．     

燃煤电厂锅炉PM2.5现场实测与排放特征研究

采用自设固定源PM_2.5稀释采集系统对陕西省关中地区多个燃煤电厂烟气中的颗粒物开展了现场实测与样品化学源组分分析。结果表明：4个燃煤电厂PM_2.5、PM₁₀占颗粒物比重差异较大（PM_2.5占比范围为6.92%~54.34%,PM₁₀占比范围为36.36%~73.02%）;4个燃煤电厂数浓度水平最高值（2.0×10⁴~4.0×10⁵个/cm³）差异较大,相差约1个数量级;燃煤电厂PM_2.5小粒径段颗粒物质量浓度占比大多较低,但其对PM_2.5数浓度贡献却很大（最小值>95%）,大粒径段颗粒物则相反;煤粉炉PM₁/PM_2.5较大,占比范围为35.29%~51.35%,循环流化床锅炉PM₁/PM_2.5较小,占比范围为16.62%~21.47%;SO₄^2-是燃煤电厂PM_2.5中最丰富的离子成分,各电厂SO₄^2-占总离子浓度比重范围为50.02~65.52%,Na⁺和Ca²⁺位于第2或第3位;各燃煤电厂主要检出无机元素是Si、Al、Ca、Na、Fe、Na等地壳元素;燃煤电厂PM_2.5、PM₁₀和颗粒物的排放因子范围分别为0.001~0.028 kg/t、0.002~0.086 kg/t、0.003~0.236 kg/t;除尘设施组合越复杂先进,排放因子就可能越小。     

Study on young panicle culturein vitro from wild rice of different genomes

Researches have been made on young panicle culturein vitro from wild rice of different genomes. Main results are as follows: 1. The induction frequencies of young panicle culturedin vitro from wild rice varied largely a relation to its genome. The optimal induction period of callus is the stamen and pistil differentiation stage of young panicle development. 2. Plantlets were regenerated through two ways: first, culture method, the induced calli were transferred onto differentiation medium; second, regenerate plantlets directly from young panicles of wild rice that were cultured on the differentiation medium. 3. The regeneration rate of green plantlets that obtained through cryopreservated calli inO. meyeriana was 10 times higher than that of control. Foundation item: Supported by the National Natural Science Foundation of China (39279436) Biography: SHU Li-hui(1937-), Professor, Research in plant tissue culture.