八倍体小黑麦耐盐细胞系产生的遗传机制

实验选用来源于八倍体小黑麦（×ＴｒｉｔｉｃｏｓｅｃａｌｅＷｉｔｍａｃｋ）杂种一代Ｂ、Ｃ和原种Ｆ三个品系的成对单倍体和双倍体细胞系,在１％、１．５％和２％ＮａＣｌ水平进行耐盐变异体筛选及其发生机制研究。实验和分析结果表明,未经筛选的细胞系由三类细胞组成,第一类是占大多数没有任何适盐性的细胞,第二类是具有一定适盐性的非变异体细胞,可在继代筛选中被淘汰,第三类细胞是变异体细胞;第二类细胞在杂种材料细胞系中所占比例少于原种细胞系,故杂种细胞较易筛选到耐盐变异体;比较三个级别筛选的变异体频率发现,后一级筛选的变异必须以前一级筛选的变异为基础,即耐盐变异体发生的遗传机制是基因扩增;双倍体细胞中正常同源染色体对基因扩增变异的表达有干扰作用,这种干扰因材料不同在各级筛选中呈现不稳定状态。     

水稻、小麦及高粱改良体细胞无性变异体的田间试验

参与作物组织培养(TCCP)工程的科学家们田间试验鉴定了耐盐水稻、耐盐春小麦、耐酸性春小麦及抗秋粘虫高粱的体细胞无性变异体品系.此外,日本和匈牙利亦田间试验鉴定了具有几种改良农艺性状的组培衍生水稻品系.在菲律宾Pampanga,TCCP研究人员发现,从组织培养物再生的9个水稻品系表现出显著的盐抗性的增强(与非组培物对照相比).其中有8个品系来源于离体筛选的抗盐     

药蒲公英(Taraxacum officinale Weber)耐1.5% NaCl变异体的筛选及特性分析

为提高药蒲公英的耐盐性, 用20~30 d大小的药蒲公英叶片诱导愈伤, 获得的愈伤以NaCl作为选择因子, 用直接筛选的方法, 每3周进行一次继代培养, 经3个月继代筛选获得了耐1.5% NaCl的药蒲公英愈伤组织, 将耐1.5% NaCl的药蒲公英愈伤组织接种在分化培养基上分化出芽, 之后将再生芽转接到生根培养基中进行生根培养, 经4个月得到了12株耐1.5% NaCl的药蒲公英再生植株。与野生型相比, 耐盐植株叶片宽大、叶柄粗短、叶表面覆盖白色细毛, 根粗壮较短, 花茎中部具2 cm左右的苞叶。RAPD(Random amplified polymorphic DNA , 随机扩增的多态性DNA)和SDS-PAGE检测表明, 耐盐植株与对照植株在DNA及蛋白水平上均存在明显差异。1.5% NaCl处理后, 与普通再生植株相比, 耐盐株系的抗氧化酶活性明显提高, 脯氨酸含量上升幅度更为显著, 而丙二醛(MDA)含量降低, 其主要药用成分黄酮的含量显著增加。这些结果说明耐盐植株的抗氧化防御能力明显增强。以上结果表明耐1.5% NaCl的药蒲公英再生植株为耐1.5% NaCl药蒲公英变异体, 这些耐盐变异体有望成为抗盐耐海水蔬菜家族的新成员。同时, 这些耐盐变异体植株比普通植株具有更高的医用商业价值。耐1.5% NaCl的药蒲公英再生变异体遗传稳定性的研究正在进行中。     

药蒲公英(Taraxacum officinale Weber)耐1.5% NaCl变异体的筛选及特性分析

为提高药蒲公英的耐盐性, 用20~30 d大小的药蒲公英叶片诱导愈伤, 获得的愈伤以NaCl作为选择因子, 用直接筛选的方法, 每3周进行一次继代培养, 经3个月继代筛选获得了耐1.5% NaCl的药蒲公英愈伤组织, 将耐1.5% NaCl的药蒲公英愈伤组织接种在分化培养基上分化出芽, 之后将再生芽转接到生根培养基中进行生根培养, 经4个月得到了12株耐1.5% NaCl的药蒲公英再生植株。与野生型相比, 耐盐植株叶片宽大、叶柄粗短、叶表面覆盖白色细毛, 根粗壮较短, 花茎中部具2 cm左右的苞叶。RAPD(Random amplified polymorphic DNA , 随机扩增的多态性DNA)和SDS-PAGE检测表明, 耐盐植株与对照植株在DNA及蛋白水平上均存在明显差异。1.5% NaCl处理后, 与普通再生植株相比, 耐盐株系的抗氧化酶活性明显提高, 脯氨酸含量上升幅度更为显著, 而丙二醛(MDA)含量降低, 其主要药用成分黄酮的含量显著增加。这些结果说明耐盐植株的抗氧化防御能力明显增强。以上结果表明耐1.5% NaCl的药蒲公英再生植株为耐1.5% NaCl药蒲公英变异体, 这些耐盐变异体有望成为抗盐耐海水蔬菜家族的新成员。同时, 这些耐盐变异体植株比普通植株具有更高的医用商业价值。耐1.5% NaCl的药蒲公英再生变异体遗传稳定性的研究正在进行中。     

运用多步正选择系统筛选木槿耐盐变异体（简报）

采用多步正选择系统,建立起木槿耐盐变异体筛选体系并获得耐盐细胞系和再生植株（分化率高达78％）,分化苗能耐盐。耐盐细胞的体积减小,积累的Na 和K 含量都相对较高,而脯氨酸则与对照差异不大。     

枸杞耐盐突变体的筛选及生理生化分析

用不同剂量60Coγ对以枸杞叶为外植体诱导出的愈伤组织进行辐射处理,并将恢复增殖的愈伤组织采用逐步提高盐浓度的方法,直到筛选出耐1%的愈伤组织变异体,通过对其生理生化的分析表明:变异体在不同盐浓度胁迫下干、鲜重增长均高于对照组;MDA含量和质膜透性、O-·2与H2O2含量都低于对照组;抗氧化酶SOD、POD、CAT活性均高于对照组;对可溶性蛋白SDS电泳结果表明:耐盐变异体有22条蛋白带,而对照组有21条蛋白带,其中31.3kD、21.0kD、18.5kD带为耐盐变异体所特有,而74.1kD、52.1kD带为对照组所特有,并且73.7kD、67.9kD的蛋白耐盐变异体含量高于对照组.     

转抗盐基因中天杨新品种扦插育苗耐盐试验

利用滨海盐碱土配制营养基质,采用盆栽扦插的方法,对转抗盐基因(mtl-D基因)中天杨新品种的耐盐能力、生长表现和根系发育状况进行了对比试验研究。结果表明,随着土壤含盐量的增加,插穗存活率降低,生长量减少,仅2 g.kg-1盐度试验组生长正常,叶色浓绿,存活率一直保持在90%以上,4 g.kg-1以上盐度试验组均因盐害而陆续枯死;土壤盐害首先作用于皮部侧根,表现为新生皮部根褐变坏死,导致枝叶发黄、萎蔫,并逐渐干枯死亡;扦插育苗前期,转基因中天杨与对照的耐盐能力和生长表现并无明显差别,其原因可能与插穗中外源基因的表达过程有关。     

诱发玉米幼胚愈伤组织及其再生植株耐盐性变异的研究

本文以玉米获白×Mo17单交种的幼胚为材料。以10g/1NaCl为起始浓度,利用逐步提高NaCl浓度的方法,筛选出耐20g/1NaCl的细胞变异体,并获得再生植株。同时对影响耐盐变异系筛选、分化的 因素进行了研究。讨论了愈伤组织变异体及其再生植株的稳定性。     

'德抗961'小麦耐盐生理特性研究

以常规高产小麦品种山农215953为对照,在室内200 mmol/L NaCl胁迫条件下,对小麦品种德抗961的耐盐生理特性从水分状况维持和抗氧化保护系统两方面进行了研究.结果显示,盐胁迫条件下,德抗961较对照山农215953可保持较低的膜脂过氧化程度、较好的膜完整性以及较高的SOD、CAT、POD等抗氧化酶活性,从而保持细胞膜的有序性和稳定性;同时,德抗961通过主动积累如游离脯氨酸、可溶性糖、蛋白质以及甘氨酸甜菜碱等渗透调节物质,降低渗透势,提高渗透调节能力,从而保持相对良好的叶片水分状况.表明德抗961的渗透调节能力强及相对较高的抗氧化水平可能是其耐盐性强的主要生理因素.     

大豆耐盐相关QTLs鉴定和功能基因研究进展

大豆(Glycine max (L.) Merill)是重要的粮食作物和经济作物,盐胁迫能造成大豆产量的大幅度降低。本文综述了通过正向遗传学手段获得的大豆耐盐数量性状位点(Quantitative trait locus, QTL)以及通过反向遗传学方法获得的大豆耐盐功能基因方面的研究进展。目前,正向遗传学发掘基因主要有图位克隆(Map-based cloning)和全基因组关联分析(Genome-wide association study, GWAS)两种方案,其中通过图位克隆在大豆中已经获得了6个耐盐QTL位点并且定位了1个重要的耐盐基因;利用GWAS在大豆中获得了1个耐盐功能基因。利用反向遗传学在大豆中获得了大量的耐盐相关功能基因并在模式植物中验证了其功能,主要包括离子转运蛋白基因和转录因子基因。这些研究为揭示大豆耐盐分子机制以及通过分子标记辅助育种或转基因技术创制耐盐大豆奠定了基础。     

小麦-簇毛麦新种质山农05078的鉴定(简报)

簇毛麦（Dasypyrum villosum,VV,2n=14）属禾本科小麦族小麦亚族簇毛麦属,分蘖力比较强,多花,籽粒蛋白质含量高,耐旱,抗寒性好,高抗锈病、全蚀病和白粉病。硬簇麦（AABBVV,2n=42）是利用硬粒小麦和簇毛麦杂交人工合成的双二倍体,保留了簇毛麦的优良性状和特点,是进行小麦遗传改良的优良中间材料。人工合成小麦Am3（AABBDD,2n=42）是利用波斯小麦与粗山羊草杂交后染色体加倍而形成的双二倍体,但它所具有的A、B、D染色体组可能与经过多年分化的普通小麦所含的A、B、D染色体组不同,     

抗条锈病小偃麦双体异附加系山农87074-519的鉴定

综合利用抗性接种鉴定、细胞学分析、SSR分子标记和基因组原位杂交(GISH)技术相结合的方法,对从长穗偃麦草与小麦复合杂交后代中选育的抗条锈病种质系山农87074-519进行了鉴定。结果表明,山农87074-519的根尖细胞染色体数目2n=44,花粉母细胞减数分裂中期I(PMCMI)绝大多数细胞内可观察到22个二价体,平均染色体构型2n=44=21.82Ⅱ 0.36Ⅰ,它与普通小麦中国春杂种F1的多数花粉母细胞内染色体构型为2n=21Ⅱ 1Ⅰ,因此它是1个附加了1对长穗偃麦草染色体的双体异附加系;以假鹅冠草St基因组总DNA作探针进行原位杂交发现山农87074-519的44条染色体中有2条出现黄绿色杂交信号,且杂交信号遍布整条染色体,证明其附加的长穗偃麦草染色体为St基组;利用SSR分子标记技术,在170对SSR引物中筛选出特异引物BARC165,它能稳定地在山农87074-519中扩增出长穗偃麦草特异标记BARC165268;将长穗偃麦草中BARC165的特异扩增片段克隆测序后制备成探针进行原位杂交,可在山农87074-519的间期染色体和有丝分裂中期染色体检测到杂交信号。山农87074-519综合农艺性状较好,对条锈病免疫,其抗性基因为显性,且位于附加的长穗偃麦草St基组染色体上,暂将其表示为YrSt。该种质系在小麦的遗传改良中具有重要利用价值。     

小麦抗旱相关生理性状的QTL分析

利用RIL群体131个系,在小麦中首次对超氧化物岐化酶活性、可溶性蛋白含量、脯氨酸含量、硝酸还原酶活性、气孔导度等5个抗旱相关生理性状进行了QTL定位,共检测到超氧化物岐化酶活性、可溶性蛋白含量、硝酸还原酶活性等3个性状5个加性QTL,涉及1D、2B、5A、7B共4条染色体,可解释表型变异的8.74％～36.96％。其中．3个QTL即QSDd,sdau-7B、Qspc,sdau-1D、QNra,sdau-1D贡献率较大,分别为29.28％、23.11％和36．96％,其加性效应都源于山农483。讨论了可能用于标记辅助选择的QTL及其分子标记。     

测墒补灌对不同穗型小麦品种耗水特性和干物质积累与分配的影响

在田间试验条件下, 以中穗型小麦(Triticum aestivum)品种‘山农15’和大穗型品种‘山农8355’为供试材料, 设置3个0-140 cm土层土壤相对含水量处理: W0 (拔节期65%, 开花期60%)、W1 (拔节期70%, 开花期70%)、W2 (拔节后8天70%, 开花后8天70%), 采用测墒补灌的方法补充土壤水分达到目标相对含水量, 对两个不同穗型小麦品种的耗水特性和干物质积累与分配进行了研究。结果表明: (1)两品种籽粒产量均以W0处理最低, ‘山农15’ W1和W2处理无显著差异, ‘山农8355’ W1处理显著高于W2处理; 两品种W1处理的水分利用效率和灌溉水利用效率均显著高于W2处理。‘山农15’ W1处理的籽粒产量和灌溉水利用效率分别显著低于和高于‘山农8355’的W1处理, 水分利用效率无显著差异; 两品种W2处理的籽粒产量、水分利用效率和灌溉水利用效率均无显著差异。(2)两品种总耗水量以W0处理最低, ‘山农15’ W1处理显著低于W2处理, ‘山农8355’两处理无显著差异; 两品种W1处理的土壤供水量及其占总耗水量的比例显著高于W2处理。‘山农15’ W1处理的总耗水量和灌水量占总耗水量的比例显著低于‘山农8355’, 土壤供水量占总耗水量的比例显著高于‘山农8355’; 两品种W2处理总耗水量, 土壤供水量及其占总耗水量的比例无显著差异。(3)两品种W1处理成熟期干物质积累量显著高于其他处理, W1处理提高了‘山农8355’开花后干物质积累量及其对籽粒的贡献率, 对‘山农15’无显著影响。‘山农15’ W1和W2处理成熟期干物质积累量显著低于‘山农8355’, 开花前贮藏同化物向籽粒的转运量和转运率、对籽粒的贡献率均显著高于‘山农8355’, 开花后干物质积累量及其对籽粒的贡献率低于‘山农8355’。综合考虑干物质积累与分配、籽粒产量、水分利用效率和灌溉水利用效率, W1处理是两品种节水高产的最佳土壤相对含水量处理。     

小黑麦×小滨麦后代1RS·1BL易位系的选育和鉴定

利用细胞学、染色体C分带和原位杂交方法,对八倍体小黑麦劲松49与八倍体小滨麦杂种后代选育的稳定遗传材料山农030-1进行了鉴定。染色体观察结果表明,山农030-1根尖染色体数目为42,花粉母细胞减数分裂中期I(PMC MI)染色体构型为2ｎ=21Ⅱ。分别以黑麦、滨麦草基因组DNA为探针进行原位杂交,表明山农030-1含有一对黑麦和小麦的整臂易位染色体,不含有来自滨麦草的遗传物质。染色体C分带结果进一步表明此材料为1RS·1BL易位系。接种鉴定表明,山农030-1对白粉病免疫。Abstract: In the progenies of octoploid Triticale Jinsong49 crossed with octoploid Tritileymus, a stable germplasm line named Shannong030-1 was obtained. Its chromosome number was 42, and 21 bivalents could be observed at PMC MI. FISH analysis by using genomic DNAs from S. cereale and genomic DNAs from L. mollis as probes,respectively, showed that Shannong030-1 contained a pair of translocation chromosome between rye and wheat, and had no genetic materials from L. mollis . C-banding analysis showed that it was translocation lines of 1RS·1BL.Resistance identification showed Shannong030-1 was immune to powdery mildew.     

Transthyretin Ser 6 gene frequency in individuals without amyloidosis

Transthyretin (TTR) Ser 6 was originally described in a Scottish kindred without amyloidosis. This variant, arising from a GA transition in codon 6 that destroys an MspI site and creates a BsrI site, was present in none of 50 controls, and was therefore throught to be rare. This variant has subsequently been found in a normal human cDNA liver library and in two unrelated patients with familial amyloidosis and other TTR variants, raising the question whether it is actually a common polymorphism. To address this question, we performed PCR and restriction digestion of 574 DNA samples from people without evidence of amyloidosis or a known family history of amyloidosis. The TTR Ser 6 allele frequency was 33/558 (.060) in Caucasians (including 8/192 (.04)) in North American Ashkenazic Jews, 16/218 (.07) in North American non-Jews, and 9/148 (.06) in Portuguese), 3(242 (.01) in African Americans, 0/140 in Africans, and 0/208 in Asians. These data are most suggestive of a single Caucasian founder and the known 25% admixture of Caucasian genes in the African-American population. Alternatively, as this variant arose from a transition at a CG dinucleotide hot spot, it may have arisen on multiple occasions. These data indicate that TTR Ser 6 is a common non-amyloidogenic population polymorphism in Caucasians.     

Postanthesis allocation of photosynthates and grain growth in wheat cultivars as affected by source/sink change

Two wheat cultivars, Hesheng 2 with large grain yield potential, and Shannong 505 with small grain yield potential, were used for investigating the responses of postanthesis photosynthesis, dry matter accumulation and allocation, and grain growth to source/sink changes. At the initial grain filling stage, Hesheng 2 was sensitive to source reduction leading to an increase of net photosynthetic rate (P_N) by 10 %; however, little effect of sink reduction was observed. In Shannong 505, P_N was obviously decreased by sink reduction, and changed a little after source reduction. At the rapid grain filling stage, Hesheng 2 was sensitive to both source and sink reduction resulting in the increase or decrease of P_N, respectively. However, the response of P_N in Shannong 505 to source/sink changes was similar to that in previous stage. The dry matter (DM) accumulation after anthesis was affected by source/sink changes. In Hesheng 2, the decrease in DM was higher than that in Shannong 505 after the same source or sink reduction. Source reduction caused a decrease in the allocation of DM to the sheath and stem, and promoted the reserve photosynthates to be reallocated to grain. The effect of sink reduction was contrary. The grain mass of Hesheng 2 was more easily regulated by source/sink changes than that of Shannong 505. The effect source/sink changes on grain mass was in order upper > basal > middle spikelets on spike. As for a spikelet, the effect was found mainly in the grain mass at the positions 3 and 4 from base of the spikelet. This revised version was published online in July 2006 with corrections to the Cover Date.     

Crystal Morphology Engineering of Pharmaceutical Solids: Tabletting Performance Enhancement

Crystal morphology engineering of a macrolide antibiotic, erythromycin A dihydrate, was investigated as a tool for tailoring tabletting performance of pharmaceutical solids. Crystal habit modification was induced by using a common pharmaceutical excipient, hydroxypropyl cellulose, as an additive during crystallization from solution. Observed morphology of the crystals was compared with the predicted Bravais–Friedel–Donnay–Harker morphology. An analysis of the molecular arrangements along the three dominant crystal faces [(002), (011), and (101)] was carried out using molecular simulation and thus the nature of the host–additive interactions was deduced. The crystals with modified habit showed improved compaction properties as compared with those of unmodified crystals. Overall, the results of this study proved that crystal morphology engineering is a valuable tool for enhancing tabletting properties of active pharmaceutical ingredients and thus of utmost practical value.     

Differences in physiological characteristics between two wheat cultivars exposed to field water deficit conditions

We investigated various physiological characteristics of two wheat (Triticum aestivum L.) cultivars differing in drought tolerance, i.e., Shannong16 (a drought-tolerant cultivar) and Weimai8 (a high-yield wheat cultivar under well-watered conditions), under field drought conditions. The experiments were conducted over a two-year period. Drought stress (DS) was imposed by controlling irrigation and sheltering the plants from rain. Compared with Weimai8, Shannong16 exhibited the better water balance, the higher osmotic adjustment, the slower degradation of chlorophyll, and the higher net photosynthetic rate under drought-stress conditions. At the same time, we observed that Shannong16 maintained more integrated chloroplast and thylakoid ultrastructure in flag leaves than Weimai8 under field drought stress. The different levels of antioxidant competence, indicated by MDA content, antioxidant enzyme activities, and the level of superoxide radicals observed in the two wheat cultivars may be involved in the different levels of drought resistance of these cultivars.     

Enrichment of a common wheat genetic map and QTL mapping for fatty acid content in grain

DArT and SSR markers were used to saturate and improve a previous genetic map of RILs derived from the cross Chuan35050 × Shannong483. The new map comprised 719 loci, 561 of which were located on specific chromosomes, giving a total map length of 4008.4 cM; the rest 158 loci were mapped to the most likely intervals. The average chromosome length was 190.9 cM and the marker density was 7.15 cM per marker interval. Among the 719 loci, the majority of marker loci were DArTs (361); the rest included 170 SSRs, 100 EST-SSRs, and 88 other molecular and biochemical loci. QTL mapping for fatty acid content in wheat grain was conducted in this study. Forty QTLs were detected in different environments, with single QTL explaining 3.6-58.1% of the phenotypic variations. These QTLs were distributed on 16 chromosomes. Twenty-two QTLs showed positive additive effects, with Chuan35050 increasing the QTL effects, whereas 18 QTLs were negative with increasing effects from Shannong483. Six sets of co-located QTLs for different traits occurred on chromosomes 1B, 1D, 2D, 5D, and 6B.