Generation of the genetic mutant population for the screening and characterization of the mutants in response to drought in maize

The autonomous Mutator(Mu)transposon in maize(Zea mays L.)lines 115F,330I,and 715D were crossed with inbred lines B73,Mo17,97108,and H9-21,M1were self-pollinated to establish the Mu insertion-mutagenized M2seeds pool and The M2plants encompassed a large amount of biological variation relevant to improving agronomy traits by observed in the field.Next we statistically analyzed these candidate mutants.Under drought stress,we screened seedlings at 3-leaf stage by using the infrared thermography,and successfully got 108 droughtinsensitive and 121 drought-sensitive candidate mutants from more than 38,000 lines,which temperature were significant different with others.These candidate mutants were primarily analyzed by infrared thermography,chlorophyll fluorescence,leaf water losing,photosynthetic characteristics,content of soluble sugars,soluble protein,and proline assay.The selected mutants were cloned by MuTAIL–PCR methods.Here we provide the better genetic materials for research on maize breeding for drought tolerance.     

盐胁迫对主要造林树种种子活力及幼苗生理特性的影响

在水培条件下进行刺槐（Ｒｏｂｉｎｉａｐｓｅｕｄｏａｃａｃｉａ）、马尾松（Ｐｉｎｕｓｍａｓｏｎｉａｎａ）、水杉（Ｍｅｔａｓｅｑｕｏｉａｇｌｙｐｔｏｓｔｒｏｂｏｉｄｅｓ）种子和幼苗的耐盐性试验。盐浓度为０．０％,０．１％,０．２％,０．３％,同时用２５０×１０－６多效唑（ＭＥＴ）测定预处理种子后的耐盐性。结果表明：盐浓度越大,种子发芽势及发芽率越低,简化苗木活力指数（ＳＶＩＳ）越小;ＭＥＴ对种子发芽势、发芽率及ＳＶＩＳ有抑制作用。但盐浓度越大,种子发芽率、ＳＶＩＳ越大;根茎比随盐浓度的增加而降低,ＭＥＴ有增加根茎比值的趋势;幼苗体内游离脯氨酸含量随盐浓度增加而增加,ＭＥＴ能提高脯氨酸的含量;在试验盐浓度下,刺槐、马尾松耐盐能力为０．２％～０．３％,水杉０．１％～０．２％。在沙培条件下,用２５０×１０－６硼酸,０．３％氯化钙和２５０×１０－６ＭＥＴ处理刺槐、湿地松（Ｐｉｎｕｓｅｌｉｏｔｉｉ）、侧柏（Ｐｌａｔｙｃｌａｄｕｓｏｒｉｅｎｔａｌｉｓ）种子,其耐盐能力增强。其中,氯化钙处理湿地松效果较好;硼酸处理侧柏效果好,ＭＥＴ处理居中。３种处理对刺槐的耐盐能力影响不大。     

Transpositional behaviour of the Ds element in the Ac/Ds system in rice

Insertional mutagenesis based on maize Activator/Dissociator (Ac/Ds) transposons is becoming a ma- jor approach used to produce a saturated mutant collection in rice. In this research, Ds-T-DNA trans- formed homozygotes were crossed with Ac-T-DNA transformed homozygotes in order to establish an Ac/Ds transposon system in rice. The successive investigation of Ds transposition from F1 to F5 gen- erations indicated that the frequencies of germinal transposition increased over successive genera- tions and reached 54.2% in F3 generation. The Ds transposition pattern revealed that a Ds transposition induced an approximately 170-bp deletion of T-DNA sequence and another Ds transposition carried a 272-bp T-DNA sequence. Using thermal asymmetric interlaced PCR (TAIL-PCR), some flanking se- quences of the Ds element were amplified. Analyses of 17 Ds-flanking sequences showed that five Ds were inserted into gene regions. The Ds could transpose not only to the linked sites but also to the unlinked sites. The frequency of inter-chromosomal transposition of Ds was 33.3%.     

Accurate localization and excision of genomic islands in four strains of <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> and <Emphasis Type="Italic">Pseudomonas fluorescens</Emphasis>

Mobile genomic islands (GIs) can be excised from the chromosome, then form a circular intermediate and be reintegrated into the chromosome by the GI internal integrase. Some mobile GIs can also be transferred into a new receptor cell by transformation, conjugation, or transduction. The action sites of the integrase are usually flanked direct repeats (DRs) of the GIs. Accurate localization of the flanking sequences is a precondition for determining the mobility of the GI. Mobile GIs are generally associated with transfer RNAs (tRNAs). Based on the correlation between flanking sequences and tRNA sequences, the flanking sequences of 11 putative mobile GIs in Pseudomonas aeruginosa PAO1, P. aeruginosa PA14, P. fluorescens Pf-5 and P. fluorescens Pf0-1 were identified. Among the 11 GIs, Pf0-1GI-1 is responsible for benzoate degradation. PAO1GI-1, Pf5GI-2, Pf5GI-3, and Pf5GI-4 were confirmed experimentally to be excised from a chromosome to form a circular intermediate. The action sites of the integrases are these GIs direct repeats. Due to distinct DRs, cutting sites for the internal integrase of PAO1GI-1, Pf5GI-2, Pf5GI-3 and Pf5GI-4 were determined outside the T-loop of the tRNA^Gly gene, outside the anticodon loop of the tRNA^Ser gene and tRNA^Lys gene, and at the asymmetric 3′-end of the tRNA^Leu gene, respectively. PAO1GI-1 and other mobile GIs may be transferred into many different strains that belong to different phyla because of the clear flanking sequences. This study describes basic information about the action sites of the integrases, assesses the mobility of GIs, and can help design and transfer mobile GIs to candidate strains.     

Characterization, development and exploitation of EST-derived microsatellites in Gossypium raimondii Ulbrich

COTTON IS AN IMPORTANT GLOBAL CASH CROP. IN THE RECENTYEARS, MOLECULAR MARKER TECHNOLOGY HAS BEEN WIDELY APPLIED TO SUCH STUDIES ON COTTON AS GENETIC MAP- PING[1―4], VARIETY PURITY DETECTION[5], GENETIC DIVERSITY ANALYSIS[6], MOLECULAR MARKER-ASSISTED BR…     

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     

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…     

A miniature insertion element transposable in Microcystis sp. FACHB 854

A 186-bp sequence with imperfect terminal inverted repeats and target direct repeats but without any transposase-encoding capacity was found to be transposable in an isolate derived from Microcystis sp. FACHB 854. This miniature insertion element, designated as ISM854-1, and with its homologues present at least 10 copies in the genome of Microcystis FACHB 854, is inserted into the 8-bp long and AT-rich target sequences, but none or few in other Microcystis strains. A variant of ISM854-1, denoted ISM854-1A, has perfect inverted repeat sequences and may transpose in pairs in a structure like a composite transposon. This is the first report of non-autonomous transposition of a mini-IS in a cyanobacterium.     

玉米幼苗热激诱导抗冷性过程中钙的效应

 玉米幼苗在冷胁迫前经过热激处理或CaCl₂浸种后再热激处理,其存活率,抗氧化酶GR的活性、可溶性蛋白质中热稳定蛋白质和游离脯氨酸的含量在的冷胁迫中均发生了变化,发现热激处理幼苗的这些参数高于对照,而最高的是CaCl₂浸种后再热激处理的,表明热激能提高玉米幼苗的抗冷性,而Ca²⁺对上述热激处理有加强作用.     

广西北海大冠沙白骨壤树上大型固着动物的数量及其分布

本文研究了广西北海大冠沙白骨壤树上大型固着动物的种类组成、数量分布特点及其对植物体的危害。结果表明,大冠沙白骨壤群落中的大型固着生物有10种,分属3门4纲6科,其中在白骨壤植株上的主要种类为白条小藤壶、潮间藤壶、褶牡蛎和黑荞麦蛤4种。危謇程度最大的是白条小藤壶,其湿重生物量占总湿重生物量的百分比高达72.3%～100%。受害程度最大的是向海林带的枝和叶,其动物生物量干重与植物生物量干重的比值(a/p)可达30.49×10~(-2)和30.43×10~(-2)。就整个红树林区而言,危害程度为向海林带(21.68×10~(-2))>中间林带(7.88×10~(-2))>向陆林带(0.01×10~(-2)).同一林带的植株受害程度为较低树层的大于较高树层的.密度和生物量的分布也与此相类似.     

Constitutive activation of AtMEK5, a MAPK kinase, induces salicylic acidindependent cell death in Arabidopsis thaliana

In most circumstances, plants are able to defendthemselves against pathogen attack. The mechanisms ofplants resisting the invasion of pathogens often includeinduction of hypersensitive response (HR) cell death, theactivation of defense genes and the production of antim-icrobial phytoalexins[1—3]. Plant defense responses are ini-tiated either by the interaction between a plant resistance(R) gene and a pathogen avirulence (avr) gene or by thebinding of a non-race-specific elicitor to its recep…     

褐家鼠和黄胸鼠取食行为研究

在４５ｍ２室内人工模拟自然条件下进行褐家鼠和黄胸鼠取食行为生态学特征的研究。观察结果显示，两种鼠对所提供的食物的日平均取食量和觅食时所付出的活动行为强度均无明显差异，但在日内的７时、１５时和２３时之间的摄食量和活动量的差异均极显著。当所提供的食物源有选择机会时，褐家鼠选择的平均取食量是玉米碎＞稻谷＞大米；黄胸鼠是玉米碎＞大米＞稻谷，无食物选择机会时褐家鼠的取食量是稻谷＞玉米碎＞大米；黄胸鼠是大米＞玉米碎＞稻谷。褐家鼠和黄胸鼠在寻找选择食物的过程中，通常付出的活动行为强度（量）是２３时点＞７时点＞１５时点。褐家鼠和黄胸鼠的取食行为特征表现可用“摄食时间＋食物源＋行为反应”的模式表示。同时指出鼠的取食行为与防治实践之间关系的重要性。     

黑老虎植株不同部位木脂素和萜类成分分析

黑老虎(Kadsura coccinea)为传统中药材，为深入了解木脂素类和萜类成分在植株不同部位的分布和积累状况，采用广泛靶向代谢组学技术鉴定其叶、茎、根中的代谢物，基于结构配置解析木脂素和萜类的多样性和丰度。结果显示：在黑老虎植株中共鉴定出39个木脂素类化合物，根(32个，特有13个)>茎和叶(同为24个)，3个部位含17个相同的木脂素类化合物；同时，鉴定出41个萜类成分，根(39个)>茎和叶(同为36个)，3个部位含34个相同的萜类化合物。黑老虎根部木脂素类化合物丰度(114.62×10⁶)远高于叶(23.37×10⁶)和茎(25.96×10⁶),萜类化合物丰度为根(74.16×10⁶)>叶(37.06×10⁶)>茎(17.36×10⁶),根部与叶、茎相同及特有的木脂素和萜类化合物丰度均较高。日本南五味子木脂素G、乙酰表戈米辛R、Kadcoccilactone F等在黑老虎根中高度富集，在叶、茎、根中都鉴定出Kadcoccilactone Q-R、Kadcoccilactone M等三萜类化合物，表明黑老虎具有较高的研究与利用价值。     

Construction of cDNA library from iron-deficiency induced maize roots and screening and identification of iron-stress geneFdr3

To isolate Fe-deficient related (Fdr) genes, an expression cDNA library of 4.5×10⁵ pfu/μg has been constructed from maize roots in iron-stress. 6 clones have been screened from the cDNA library by differential hybridization screening. It is proved that anFdr3 cDNA clone expressed stronger under iron-deficient condition than under iron-sufficient one by Northern blot and Western blot.     

Genetic transformation of calli from maize and regeneration of herbicide-resistant plantlets

The herbicide-resistant geneals ofArabidopsis thaliana has been transferred into embryonic calli of maize by microprojectile bombardment. We have obtained chlorsulfuron-resistant calli and regenerated plantlets through selection by herbicide chlorsulfuron. The results of PCR analysis and Southern blotting indicate that geneals has been transferred into some plantlets. The test of spraying chlorsulfuron indicated that the transgenic plantlets had favorable herbicide-resistant trait. The purpose of the research was to obtain chlorsulfuron-resistant transgenic maize and hope that this kind of high efficient herbicide could be widely used in rotation soil of wheat and maize. In addition, through spraying herbicide, we could eliminate the hybrid plants and thereby increase the purity of F1 seeds.     

Transposition of Tn554 does not generate a target duplication

Transposable elements from prokaryotic and eukaryotic organisms are discrete DNA segments bounded by inverted or directly repeated sequences that insert into non-homologous DNA in a reaction that is independent of the general recombination functions of the host. The mechanisms proposed generally involve a staggered double-stranded scission of the target DNA, ligation to the nicked ends of the transposable element, and replication of the element, resulting in the generation of a directly repeated oligonucleotide target sequence flanking the new copy of the element. Most transposons have a relatively low degree of target site specificity coupled with a low insertion frequency. Tn554, a Staphylococcus aureus transposon which specifies resistances to erythromycin and spectinomycin, displays an unusually high degree of insertion specificity. Tn554 transposes with high efficiency to a unique ('primary') site in the S. aureus chromosome and only rarely (less than 10(-6) per transductant) to other, secondary sites. We report here the nucleotide sequences surrounding the junctions of Tn554 in three independent 'primary' insertions and two 'secondary' insertions of the transposon. Two unusual features are revealed: first, the termini of Tn554 contain neither inverted nor directly repeated sequences. Second, transposition of Tn554 does not generate the short direct repeats of the target DNA that are characteristic of other transposable elements. These results suggest that the mechanism of Tn554 insertion may be significantly different from that of other transposons.     

潮间带贝克喜盐草土壤种子库空间分布及其生物影响因素

【目的】为探讨贝克喜盐草(Halophila beccarii Ascherson)种群更新及濒危机制,研究其土壤种子库空间分布特征及其生物影响因素。【方法】对广西合浦榕根山、防城港交东、防城港山心3个贝克喜盐草典型分布点进行调查取样,统计(或称量)样品种子、直立茎、雌花、果实数量、地上部分生物量和地下部分生物量。【结果】从水平方向分布来看,相同方向上不同距离处贝克喜盐草土壤种子库密度差异不显著,海草场潮沟、外海、海岸方向一侧的土壤种子库密度在榕根山和交东分别为271粒/m~2、180粒/m~2、52粒/m~2和52粒/m~2、23粒/m~2、16粒/m~2,差异显著,可见该海草种子水平空间分布格局受到落潮水流与落潮后流向潮沟的微水流的影响,受距离影响较弱;垂直方向上,上层(0~4cm)、中层(4~8cm)、下层(8~12cm)土壤底质中贝克喜盐草种子数量分别占总量的73.0%、20.8%和6.2%,上层土壤中种子数量显著高于中层和下层(P0.05),表明绝大部分的种子分布于土壤表层(0~8cm);贝克喜盐草海草场土壤种子库密度与其种群雌花密度、直立茎密度存在极显著线性正相关关系,其相关系数r分别为0.722 9(P0.01)、0.845 1(P0.01),与果实密度、地上部分生物量及总生物量呈多项式曲线关系。【结论】贝克喜盐草土壤种子库的水平空间分布受潮沟、入海河流等外界环境和地上植被的种群属性、种子自身特性等因素的影响,垂直方向上主要分布在表层底质中,且地上植被的种群属性是决定土壤种子库密度的关键因素。     

Comparative analysis of the dissolution kinetics of galena in binary solutions of HCl/FeCl<Subscript>3</Subscript> and HCl/H<Subscript>2</Subscript>O<Subscript>2</Subscript>

A comparative study of the dissolution kinetics of galena ore in binary solutions of FeCl₃/HCl and H₂O₂/HCl has been undertaken. The dissolution kinetics of the galena was found to depend on leachant concentration, reaction temperature, stirring speed, solid-to-liquid ratio, and particle diameter. The dissolution rate of galena ore increases with the increase of leachant concentration, reaction temperature, and stirring speed, while it decreases with the increase of solid-to-liquid ratio and particle diameter. The activation energy (E _a) of 26.5 kJ/mol was obtained for galena ore dissolution in 0.3 M FeCl₃/8.06 M HCl, and it suggests the surface diffusion model for the leaching reaction, while the E _a value of 40.6 kJ/mol was obtained for its dissolution in 8.06 M H₂O₂/8.06 M HCl, which suggests the surface chemical reaction model for the leaching reaction. Furthermore, the linear relationship between rate constants and the reciprocal of particle radius supports the fact that dissolution is controlled by the surface reaction in the two cases. Finally, the rate of reaction based on the reaction-controlled process has been described by a semiempirical mathematical model. The Arrhenius and reaction constants of 11.023 s⁻¹, 1.25×10⁴ and 3.65×10² s⁻¹, 8.02×10⁶ were calculated for the 0.3 M FeCl₃/8.06 M HCl and 8.06 M H₂O₂/8.06 M HCl binary solutions, respectively.     

Iterative Method for Constructing Complete Complementary Sequences with Lengths of 2^mN

Complete complementary sequences are widely used in spectrum spread communications because of their ideal correlation functions. A previous method generates complete complementary sequences with lengths of N^nN （n,N ∈ Z^＋）. This paper presents a new iterative method to construct complete complementary sequences with lengths of 2^mN （m,N ∈ Z^＋）. The analysis proves that this method can produce many sequence sets that do not appear in sequence sets generated by the former method, especially shorter sequence sets. The result will certainly increase the application of complete complementary sequences in communication engineering and related fields.     

Ubi1 intron-mediated enhancement of the expression of Bt cry1Ah gene in transgenic maize （Zea mays L.）

The cry1Ah gene was one of novel insecticidal genes cloned from Bacillus thuringiensis isolate BT8. Two plant expression vectors containing cry1Ah gene were constructed. The first intron of maize ubiqutinl gene was inserted between the maize Ubiquitin promoter and cry1Ah gene in one of the plant expressing vectors （pUUOAH）. The two vectors were introduced into maize immature embryonic calli by microprojectile bombardment, and the reproductively plants were acquired. PCR and Southern blot analysis showed that foreign genes had been integrated into maize genome and inherited to the next generation stably. The ELISA assay to T1 and T2 generation plants showed that the expression of CrylAh protein in the construct containing the ubil intron （pUUOAH） was 20% higher than that of the intronless construct （pUOAH）. Bioassay results showed that the transgenic maize harboring cry1Ah gene had high resistance to the Asian corn borers and the insecticidal activity of the transgenic maize containing the ubil intron was higher than that of the intronless construct. These results indicated that the maize ubil intron can enhance the expression of the Bt cry1Ah gene in transgenic maize efficiently