小麦抗白粉病新基因PmCD1和抗条锈病基因YrCD的鉴定及分子标记筛选

本研究对自育小麦白粉病抗源“07鉴126”和条锈病抗源CD1437、CD0534-5进行抗性遗传分析和微卫星引物的筛选，建立了与PmCD1和YrCD抗病基因连锁的SSR分子标记，主要研究结果如下： 1.小麦白粉抗源“07鉴126”抗白粉病基因的鉴定和分子标记的建立 品系“07鉴126”对我国目前白粉菌强优势生理小种E09、E11和其它多种小种表现免疫或高度抵抗。Pm-sus是07鉴126的自然突变感病株。利用“07鉴126”和Pm-sus的F2抗病性分离群体进行抗条锈病性遗传分析和分子标记定位，结果表明，“07鉴126”的白粉抗性为显性单基因控制的全生育期抗性，暂命名为PmCD1；并筛选到了与PmCD1共分离的显性SSR分子标记Xbarc183。系谱分析和分子标记分析表明PmCD1来源于荆州黑麦。抗谱分析表明PmCD1不同于已知的黑麦抗白粉基因，是一个新的抗白粉病基因。Xbarc183这一分子标记的建立为PmCD1的分子标记辅助选择和抗病基因累加提供了方便。 2.小麦条锈抗源CD1437抗条锈病基因的鉴定和分子标记的建立 利用对优势条锈菌小种条中32免疫的小麦品系CD1437及其自然突变感病株Yr-sus杂交构建F2、F3抗病性分离群体。抗条锈病性遗传分析结果显示，1437的抗条锈性为显性单基因控制的全生育期抗性，该基因暂命名为YrCD。SSR分析发现，位于1B染色体上的7个SSR标记Xcfd65、Xgwm11、Xgwm18、Xbarc187、Xwmc406、 Xwmc419、Xwmc216依次分布在YrCD的一侧，与YrCD的遗传距离在1.7 cM至9.2 cM。YrCD和YrCH42的等位性分析显示二者可能为等位基因。YrCD和Yr24、Yr26的抗谱相似。系谱分析和分子标记分析表明贵农20是YrCD的供体。本研究推测YrCD、Yr24、Yr26和YrCH42可能是等位基因，并推测Yr-sus是缺失突变体。 3. 小麦条锈抗源CD0534-5抗条锈病基因的鉴定 利用对条中32免疫的小麦抗条锈病品系CD0534-5及其感病重组自交系CD0534-4建立F2抗病性分离群体。抗条锈病性遗传分析表明，CD0534-5的条锈抗性由两对独立的显性主效基因控制。用BSK法分析，发现其中一对基因与SSR分子标记Xgwm11、Xgwm18、Xwmc128、Xwmc419连锁，该基因是来源于贵农20的YrCD。另一抗性基因来源贵农19，是极有利用价值的未知抗性基因。 This study focused on the investigation and identification of a novel powdery mildew resistant gene PmCD1 in wheat lines 07jian126 and stripe rust resistant gene YrCD in wheat lines CD1482 and CD0534-5, and screened SSR markers tightly linked to them. The main results were as follows: 1.Identification and SSR markers screening of a novel powdery mildew PmCD1 in wheat line 07jian126. Using a Pm resistant wheat line 07jian126 and its Pm susceptible mutant, a F2 population was constructed. Pedigree and genetic analyses indicated that the Pm resistance in 07jian126 was tranderred from rye (Secale cereale L.) cv. Jinzhou and was controlled by a single dominant gene. Differential test using 21 Bgt isolates revealed that the Pm resistant gene in 07jian126 is novel and was temporarily designated as PmCD1. A dominant SSR marker Xbarc183/130 bp was found co-segregated with PmCD1 in the F2 population. The diagnostic band of Xbarc183/130 bp co-segregating with PmCD1 could be used as an ideal marker in marker-assisted-selection during wheat breeding program. 2. Identification and SSR markers mapping of yellow rust resistant gene YrCD in wheat line CD1437. Wheat line CD1437 was highly resistant to predominant Chinese stripe rust race CYR32 at both seedling and adult stages. A F2 population was developed from the cross of CD1437 and its Yr susceptible mutant Yr-sus. Genetic analysis indicated line CD1437 contains a single dominant gene, temporarily designated YrCD. Seven SSR markers on the chromosome 1BS including Xcfd65, Xgwm11, Xgwm18, Xbarc187, wmc406, Xwmc419and Xwmc216 were close linked to YrCD with a genetic dsitance 1.7 cM to 9.2 cM. YrCD came from wheat cultivar Guinong 20. Allelic test of CD1437 and Chinese cultivar Chuanmai 42 indicated that YrCD and YrCH42 were allelic. Reaction patterns of YrCD and Yr24, Yr26 to 21 PST isolates were the same. These results suggested that YrCD and Yr24, Yr26, YrCH42 might be allelic. 3.Detection and identification of yellow rust resistance genes in wheat line CD0534-5 Wheat line CD0534-5 was highly resistant to predominant Chinese stripe rust race CYR32, while its recombinant inbred line CD0534-4 was susceptible. Genetic analysis with a F2 population developed from the cross of CD0534-5 and CD0534-4 indicated line CD0534-5 contains two independent dominant genes. Four SSR markers on the chromosome 1BS including Xgwm11, Xgwm18, Xwmc128, Xwmc419 were found to linked with one gene in CD0534-5. According the locations of makers and pedigree, this gene in CD0534-5 was YrCD, from cultivar Guinong 20. Another resistant gene was from cultivar Guinong 19, different with those genes on 1B such as Yr10, Yr15, Yr5 etc, was a valuable resistant gene in wheat breeding.

在盐胁迫下外源维生素浸种对小麦发芽及幼苗生长的影响

维生素(Vitamin)又称维他命，为“万年青”产品，是维持人体生命健康必需的一类低分子有机化合物质。维生素对人体健康的作用人们研究很多, 维生素可以增强人体对感染的抵抗力，降低出生缺陷及降低癌症和心脏病发病率等，一旦缺乏，肌体代谢就会失去平衡，免疫力下降，各种疾病，病毒就会趁虚而入；而维生素对作物影响的研究却很少。目前为止，尚无对用维生素浸种的方法来研究外源维生素是否对小麦种子萌发及幼苗生长起调节作用的报道，且对其在小麦抗逆性方面影响的研究甚少，对盐的胁迫抗性研究尚未有人报道。小麦（Triticum aestivum L.）属于拒盐的淡土性作物。盐害不利于小麦生长，严重影响小麦的产量和品质。本研究采用4 种不同维生素VB1、VC、VB6、VPP，分别对供试小麦品种川育12（红皮）、川育16（白皮）小麦浸种后，在一般自然条件下和逆境（盐胁迫条件）下，进行试验。探讨在正常情况下与在不同盐浓度条件下，各维生素及盐浓度对小麦发芽及幼苗生长的影响，并且比较两种不同皮色的小麦在相同盐胁迫条件下的差异表现，同时研究维生素处理的特异性，且哪种维生素对盐害缓解作用最佳。研究结果表明：在无盐胁迫（自然）条件下，对用4 种不同维生素VB1、VC、VB6、VPP 浸种小麦川育12、川育16 后的种子萌发及幼苗生长（幼苗的根长、根重、苗高、苗鲜重）的研究结果表明：4 种外源维生素浸种均对小麦发芽有调节作用，都能提高其最终发芽率。但是提高幅度有所差异。用VB6 浸种后的小麦提高幅度最多，VC 次之，VPP 提高幅度最小。同时，4 种外源维生素浸种对小麦种子的出芽速度及芽后长势也有一定的影响。VB6、VC 处理的小麦种子出芽速度最快，萌发后长势最好；VB1 出芽速度相对较慢，VPP 最慢,但都大于对照；VB1 处理长势略高于对照，VPP 处理的小麦长势则低于对照。从整体来看，VB6、VC处理促进效应明显， VB1 次之，而VPP 在某些方面无效甚至产生负效应。此外，相同的维生素处理对不同的品种的种子萌发、生长效果也存在差异，各种维生素作用于川育12 的效应均强于对川育16。进一步对幼苗根系TTC 还原力及幼苗叶片中硝酸还原酶活性进行测定、分析。研究发现：并非所有种类的维生素对幼苗根系TTC 还原力及幼苗叶片中硝酸还原酶活性的提高都有帮助。幼苗根系TTC 还原力在不同维生素处理下存在显著差异，而与小麦品种关系甚微。经VB6、VC 处理后，根系TTC 还原力测定值均显著高于对照，VB1 不明显，VPP 则略低于对照。VB6、VC 处理的幼苗叶片中硝酸还原酶的含量大于对照，VB1 与对照相差无几，而VPP 处理的川育12 幼苗叶片中的硝酸还原酶活性比对照CK 略高，而在川育16 中则略比对照CK 有所下降，呈现出抑制效应。综上结果表明：VB6、VC 具有促进种子发芽，幼苗生长及根系生长的作用，是较好的促生长剂；VPP 具有抑制作用，是较好的抑制剂，可进一步研究、开发利用。在盐胁迫条件下，对用4 种不同维生素VB1、VC、VB6、VPP 浸种川育12、川育16 后的种子萌发及幼苗生长（幼苗的根长、根重、苗高、苗鲜重）的研究结果表明：在不同盐浓度胁迫条件下， 各处理的种子萌发及幼苗生长均受到不同程度的抑制。随着盐浓度的增加, 发芽率、发芽指数和活力指数成下降趋势；幼苗的根长、根重、苗高、苗鲜重不断降低。4 种维生素处理间也表现出较大差异。VB6、VC 在每个处理中均保持对盐害的缓解作用，VB6 较VC 更易于促进发芽及幼苗生长。最终发芽率高，根系多、长、重，苗高高、重。而VB1、VPP 则表现出抑制作用。在高盐浓度150mM 时，4 种维生素浸种后的种子，其最终发芽率均不能达到40％，但VB6、VC 处理最终发芽率、苗重、根重均高于对照，VPP 最终发芽率、苗重、根重均低于对照。进一步对幼苗根系TTC 还原力及幼苗叶片中脯氨酸含量进行测定、分析。研究发现：不同盐浓度，不同维生素处理、不同品种间存在差异。随着盐浓度的增加（75mM，100mM，150mM），幼苗根系TTC 还原力活性成下降趋势，幼苗叶片中脯氨酸的积累量成上升趋势。VB6 处理脯氨酸含量增加最为明显，VC 次之，VPP 与对照接近，其变化幅度最小。经VB6、VC 处理后的幼苗根系还原强度，在不同盐浓度下，测定值均显著高于对照，VB1 不明显，VPP 则低于对照，产生负效应。此外，品种间表现不尽相同，相同的维生素处理，相同的盐浓度对不同的品种的种子萌发、生长效果也存在差异， 4 种维生素对川育16 的作用均强于川育12，但其影响趋势是一致的。说明VB6、VC 具有耐（抗）盐性，可以促进种子发芽和幼苗生长，是较好的耐（抗）盐拌种剂。 Vitamin is one kind of necessary low molecular compound for humans tosustain health and life. Lots of Studies have been done on the effectc of the vitaminsfor people. Vitamin can help people improve the body's natural resistance to disease,Drop the rate of birth defects、cacers and the incidence of the heart diseases. Ifpeople have less of them, the metabolism of the organism may throw off balance,immunity may drop off, and catch disease; Though the effects for Vitamin to thecrops are limited. up to now, there’s no one use soking seeds of wheats with vitaminsas a method, to study on how the effects will happen on the wheat seed germinationand seedling growth, and there are only few reserches on antireversion force forwheats ,none for the antireversion force in Sault stress condition.Wheat（Triticum aestivum L.）is sensitive to the salt, so the salt damage will doharm to wheat’s growth, it will have an unfavorable impact on the output and thequality of wheat.On this reaserch, we Soaking CHY12(red)、CHY16 (white) wheat seeds withVitamin C, B1, PP, B6 (50mg/L) as a pretreatment first. Then under two condition: one is in the normal environment the other is in different Salinity, we begin ourexperiments. Then disscuss on if the vitamin and salinity affect the wheat seedgermination and seedling growth, and what is the different between the two of them,the result shows that:Under the normal condition, after soaking seeds with VB1、VC、VB6、Vpp，we study on the their seed germination and the seeding growth(the root length andweights, The seedling heights and weights), it shows that all of those four kinds ofvitamin can adjust the seed germination, but different in The growth rate. VB6 isbest for increase, VC comes second,VPP is the worst. Meanwhile, those four vitaminalso have effect on the speed of the sprouting of the wheat. VB6、Vc can faster theseed germination most, and the seedlings are all doing well; VB1 do little effects onthe budding, Vpp is the worst, but all treatments are better than CK; but in Vi, VB1some what above the CK, while VPP lower than that. On the whole, the acceleratingeffect of VB6、VC are obvious, VB1 takes second place, but VPP in some aspects arenoneffective even have negative effect. Furthermore, different kind of seeds with thesame vitamin may different in seed germination and seedling growth, four vitaminson CHY16 is better than CHY12.More studies on TTC reductive capacity of roots and the activity of nitratereductase in the leaves, the reasult shows not all the vitamin can help the seedlings toimprove the TTC reductive capacity and the activity of nitrate reductase. TTCreductive capacity in different treatments shows significant differences,but notcorrelate to the variety of the wheat. The TTC reductive capacity of VB6、Vctreatments are all higher than CK, VB1 is nearly the same as CK, VPP is a littlelower than CK. Through the study of acivity of nitrate reductase, it shows that,VB6、VC are higher than CK ,VB1 is nearly the same as CK also, VPP is a little higher inthe CK of CHY12 but lower in CHY16. Through all the results above: VB6、Vc helpthe wheat seed germination, seedling growth and the growth of roots, is theperfectable factor of stimulating the growth; Vpp is a inhibition, that’ll be furtherreserch,and well develop and utilize in the future.Under the different Salinity condition, after soaking seeds with VB1、VC、VB6、Vpp，we study on the their seed germination and the seeding growth(the root lengthand weights, The seedling heights and weights), it shows that: under differentsalinity, the seed germination and the seedling growth of any treatment are inhibited.With the increase of the concentration, the germination rate, Vi、Gi all had fallen; theroot length and weight, the seedling heights and weights steadily sank down. There are also have pronounced difference between all treatments with four differentvitamins.VB6、VC in all treatments are alleviative the salt damage, VB6 is easier tocause to put forth buds than VC, and it’s quantitative value is the highest in theultimate germination rate, in root and seedlings’ hight and weight. Though the VPP、VB1 are seems to inhibite its growth. Under the high concentration150mM Nacl, theultimate germination rate in all treatments are below the 40%, but VB6、VC’squantitative values in any experiments are higher than CK,while VPP lower thanCK.Then we study on the TTC reductive capacity of roots and the content of Polinein leaves, the result shows that between the different salinity, different vitamintreatments, different varieties of the wheat have discrepancy.along with theincreasing concentraion of the salinity（75mM，100mM，150mM），TTC reductivecapacity of roots decreases, the accumulation of the content of Poline in leaves havean upward trend. The increase of VB6’s treatment are obviously, VC comessecond,VPP is nearly come up with CK, changes a little. In TTC reductive capacity of roots’s reserch, VB6、VC are higher than CK at any time,VB1 is not palpable,VPP is lower than CK, makes negative affect on wheat. In addition, varieties of thewheats are remain different, no matter it shows promoting or inhibiting, all fourvitamins have moreobvious effects on CHY16 than CHY12, but the tendency of theeffection are the same. It is say that VB6、VC can help wheat to standwith the saultwell, and promot in growth，they are the better reagent to mix with the seed.

簇毛麦（ Dasypyrum villosum）矮秆突变体的研究

株高是农作物的重要农艺性状之一，适度矮化有利于农作物的耐肥、抗倒、高产等。20世纪50年代，以日本的赤小麦为矮源的半矮秆小麦的培育和推广，使得世界粮食产量显著增长，被誉为“绿色革命”。迄今为止，已报到的麦类矮秆、半矮秆基因已达70多个，但由于某些矮源极度矮化或者矮化的同时伴随不利的农艺性状，使得真正运用于育种实践的矮源较少。因此，发掘和鉴定新的控制麦类作物株高的基因，开展株高基因定位、克隆及作用机理等方面的研究，对实现麦类作物株高的定向改良，具有重要的理论意义和应用价值。簇毛麦(Dasypyrum villosum，2n＝14，VV)是禾本科簇毛麦属一年生二倍体异花授粉植物，为栽培小麦的近缘属。本课题组在不同来源的簇毛麦杂交后代中发现了一株自然突变产生的矮秆突变体。观察分析了该突变体的生物学特性，对矮秆性状进行了遗传分析，对茎节细胞长度、花粉的活力进行了细胞学观察，考察了该突变体内源赤霉素含量及不同浓度外施赤霉素对突变体的作用，分析了赤霉素生物合成途径中的内根贝壳杉烯氧化酶（KO）和赤霉素20氧化酶(GA20ox)的转录水平，对赤霉素20氧化酶和赤霉素3-β羟化酶(GA3ox)进行了克隆和序列分析，并对GA20ox进行了原核表达和表达的组织特异性研究。主要研究结果如下：1. 该突变体与对照植株在苗期无差异，在拔节后期才表现出植株矮小，相对对照植株，节间伸长明显受到抑制，叶鞘长度基本不变。在成熟期，对照植株的平均株高为110cm，而突变株的平均株高为32cm，仅为对照植株的1/3 左右。除了株高变矮以外，在成熟后期，突变株还表现一定程度的早衰和雄性不育。I2-KI染色法观察花粉活力结果表明，对照植株花粉90%以上都是有活力的，而突变植株的花粉仅20%左右有活力。2. 突变株与对照植株的杂交F1代均表现正常株高，表明该突变性状为隐性突变。F1代植株相互授粉得到的168株F2代植株中，株高出现分离，正常株高（株高高于80cm）与矮秆植株（株高矮于40cm）的株数比为130：38，经卡方检验，其分离比符合3：1的分离比，因此推测该突变体属于单基因的隐性突变。3. 用ELISA方法检测突变株和对照植株的幼嫩种子中内源性生物活性赤霉素（GA1＋3）含量，结果表明突变株的赤霉素含量为36 ng/ml，而对照植株的赤霉素含量为900 ng/ml。对突变株外施赤霉素，发现矮秆突变株的株高和花粉育性均可得到恢复。这些结果表明该突变株为赤霉素缺陷型突变。4. 用荧光定量PCR方法比较突变株与对照植株中内根贝壳杉烯氧化酶和赤霉素20氧化酶的转录水平，结果表明突变株的KO转录水平比对照植株分别提高了6倍（苗期）和16倍（成熟期），突变株的GA20ox转录水平与对照植株在苗期无明显差异，在成熟期突变株较对照植株则提高了10倍左右。这些结果表明该矮秆突变体与赤霉素的生物合成途径密切相关，而且极有可能在赤霉素的生物合成途径早期就发生了改变。5. 以簇毛麦总基因组为模板，同源克隆了GenBank登录号为EU142950，RT-PCR分离克隆了簇毛麦的GA3ox基因cDNA全长序列，分析结果表明该cDNA全长1206bp，含完整编码区1104bp，推测该序列编码蛋白含368个氨基酸残基，分子量为40.063KD，等电点为6.27。预测的氨基酸序列含有双加氧酶的活性结构，在酶活性中心2个Fe离子结合的氨基酸残基非常保守。该序列与小麦、大麦和水稻的GA3ox基因一致性分别为98%、96%、86%。基因组序列与cDNA序列在外显子部分一致，在478-715bp和879-1019bp处分别含238bp和140bp的内含子。6. 通过RT-PCR技术克隆了簇毛麦的GA20ox基因全长，命名为DvGA20ox，GenBank登录号为EU142949。该基因全长1080个碱基，编码359个氨基酸，具有典型的植物GA20ox基因结构。该基因编码的蛋白质与小麦、大麦、黑麦草等GA20ox蛋白的同源性分别为98%，97% 和91%。该序列重组到原核表达载体pET-32a(+)上，将获得的重组子pET-32a(+)-DvGA20ox转化大肠杆菌BL21pLysS后用IPTG进行诱导表达。SDS-PAGE分析表明，DvGA20ox基因在大肠杆菌中获得了高效表达，融合蛋白分子量为55kDa。定量PCR分析表明，该基因在簇毛麦不同器官中的表达差异明显：叶片中表达水平最高，根部表达水平次之，茎部和穗中表达较弱。在外施赤霉素后，该基因的表达水平在两小时以后急剧下降，表明该基因的表达受自身的反馈调节。本研究结果认为，(1)该簇毛麦矮秆突变体为单基因的隐性突变；(2)该矮秆突变体为赤霉素敏感突变，内源赤霉素含量检测表明突变体的内源性赤霉素含量仅为对照植株的1/30；(3)荧光定量PCR结果表明突变株的赤霉素生物合成途径的关键酶基因表达水平比对照植株高，而且突变植株的赤霉素生物合成改变很可能发生在赤霉素生物合成途径的早期；(4)GA20ox有表达的组织特异性，且受到自身产物的反馈调节。 Plant height is an impotrant agronomic trait of triticeae crops.Semi-dwarf cropcultivars, including those of wheat, maize and rice, have significantly increased grainproduction that has been known as “green revolution”. The new dwarf varieties couldraise the harvest Index at the expense of straw biomass, and, at the sametime, improvelodging resistance and responsiveness to nitrogen fertilizer. Moreover, dwarf traits ofplant are crucial for elucidating mechanisms for plant growth and development aswell. In many plant species, various dwarf mutants have been isolated and theirmodles of inheritance and physiology also have been widely investigated.The causesfor their dwarf phenotypes were found to be associated with plant hormones,especially, gibberellins GAs.Dasypyrum villosum Candargy (syn.Haynaldia villosa) is a cross-pollinating,diploid (2n = 2x = 14) annual species that belongs to the tribe Triticeae. It is native toSouthern Europe and West Asia, especially the Caucasuses, and grows underconditions unfavorable to most cultivated crops. The genome of D. villosum,designated V by Sears, is considered an important donor of genes to wheat for improving powdery mildew resistance, take-all, eyespot, and plant and seed storageprotein content. A spontaneous dwarf mutant was found in D. villosum populations.The biological character and modles of inheritance of this dwarf mutant are studied.The cell length of stem cell is observed. The influence of extraneous gibberellin tothe dwarf mutant is also examined; the transcript level of key enzyme of gibberellinbiosynthesis pathway in mutant and control plants is compared. GA3ox and GA20oxare cloned and its expression pattern is researched.1. The dwarf mutant showed no difference with control plants at seedlingstage.At mature stage, the average height of control plants were 110cm and the dwarfplants were 33cm. The height of the mutant plant was only one third of the normalplants due to the shortened internodes. Cytology observation showed that theelongation of stem epidermal and the parenchyma cells were reduced. The dwarfmutant also shows partly male sterile. Pollen viability test indicates that more than80% of the pollen of the mutant is not viable.2. The inheritance modle of this dwarf mutant is studied. All The F1 plantsshowed normal phenotype indicating that the dwarfism is controlled by recessivealleles. Among the 168 F2 plants, there are 130 normal plants and 30 dwarf plants, thesegregation proportion accord with Mendel’s 3:1 segregation. We therefore proposethat this dwarf phenotype is controlled by a single recessive gene.3. Quantitative analyses of endogenous GA1+3 in the young seeds indicated thatthe content of GA1+3 was 36ng/ml in mutant plants and 900ng/ml in normal plants.The endogenous bioactive GA1+3 in mutant plants are only about 1/30 of that innormal plants. In addition, exogenously supplied GA3 could considerably restore themutant plant to normal phenotype. These results showed that this mutant wasdefective in the GA biosynthesis.4. More than ten enzymes are involved in GA biosynthesis. KO catalyzes thefirst cytochrome P450-mediated step in the gibberellin biosynthetic pathway and themutant of KO lead to a gibberellin-responsive dwarf mutant. GA20ox catalyze therate-limited steps so that their transcript level will influence the endogenous GAbiosynthesis and modifies plant architecture. The relative expression levels of genesencoding KO and GA20ox were quantified by real time PCR to assess whether thechanges in GA content correlated with the expression of GA metabolism genes andwhere the mutant occurred during the GA biosynthesis pathway. In mutant plants,the transcript levels of KO increased about 6-fold and 16-fold at the seedling stage and elongating stage respectively comparing with the normal plants. For theseedlings, there was no notable difference in the expression of GA20ox betweenmutant and normal plants. At the elongating stage, GA20ox transcript increased 10times in mutant plants, suggesting that the GA biosynthesis pathway in mutant plantshad changed from the early steps rather than the late steps.5. A full length cDNA of D. villosum gibberellin 3β-hydroxylase homology(designated as DvGA3ox) was isolated and consisted of 1206bp containing an openreading frame of 1104bp encoding 368 predicted amino acid residues. Identityanalysis showed that the gibberellin 3β-hydroxylase nucleotide sequence shared 98%,96% and 86% homology with that of wheat, barley and rice. The predicted peptidecontained the active-site Fe of known gibberellin 3β-hydroxylase and the regionhomologous to wheat, barley and Arabidopsis. The genomic clone of gibberellin3β-hydroxylase has two introns.6. The full-length cDNA of D. villosum gibberellin 20 oxidase (designated asDvGA20ox) was isolated and consisted of 1080-bp and encoded 359 amino acidresidues with a calculated mol wt of 42.46 KD. Comparative and bio-informaticsanalyses revealed that DvGA20ox had close similarity with GA20ox from otherspecies and contained a conserved LPWKET and NYYPXCQKP regions. Tissueexpression pattern analysis revealed DvGA20ox expressed in all the tissues that wereexamined and the highest expression of DvGA20ox in expanding leaves followed byroots. Heterologous expression of this cDNA clone in Escherichia coli gave a fusionprotein that about 55KD. Transcript levels of DvGA20ox dramatically reduced twohours after application of biologically active GA3, suggesting that the biosynthesis ofthis enzymes might be under feedback control.

Cytogenetic Effects of Low Doses of Energetic Carbon Ions on Rice After Exposures of Dry Seeds, Wet Seeds and Seedlings

In order to investigate the biological effects of heavy ion radiation at low closes and the different radiosensitivities of growing and non-growing plants. rice at different lift stages (dry seed, wet seed and seedling) were exposed to carbon ions at closes of 0 02, 0.2, 2 and 20 Gy. Radiobiological effects on survival, root growth and mitotic activity, as well as the induction of chromosome aberrations in root meristem. were observed The results show that radiation exposure induces a stimulatory response at lower close and an inhibitory response at higher dose on the mitotic activity of wet seeds and seedlings Cytogenetic damages are induced in both seeds and seedlings by carbon ion radiation at doses as low as 0.02 Gy Compared with seedlings. seeds are more resistant to the lethal damage and the growth rate damage by high doses of carbon ions, but are more sensitive to cytogenetic damage by low closes of irradiation Different types of radiation induced chromosome aberrations are observed between seeds and seedlings. Based on these results, the relationships between low close heavy ion-induced biological effects and the biological materials are discussed.

不同森林管理预案下友好林业局森林景观的动态变化

Knowledge about cumulative effects of forest management alternatives on forest landscape is required to make forest management decision. In this paper,a spatially explicit landscape model,LANDIS,was applied to simulate forest landscape changes in 200 years under four management alternatives (no cutting,clearcutting,selective cutting I and II) in Youhao Forestry Bureau located in Small Khingan Mountains. APACK was used to calculate distribution area of the representative species and species age cohort for six species. The results showed:1) timber harvest decreased area percentage of representative conifer species,Pinus koraiensis,Picea koraiensis and Picea jezoensis,Larix gmelinii to some extent compared to no cutting. The most influencing cutting mode for the area percentage of Tilia amurensis and Quercus mongolica was selective cutting II,followed by selective cutting I and clearcutting. To the contrast,the change of area percentage of Betula phatyphylla was contrary to the management alternatives; 2) As to species age cohort composition,timber harvest significantly changed age structure,that is,it decreased over-matured age cohort of representative species,and increased seedling and middle-age cohort (B. phatyphylla was not included,because its area percentage of over-mature age cohort was the highest under clearcutting than other three scenarios).

半湿润农田杂草及施氮对夏玉米产量及氮素利用的影响

以土垫旱耕人为土为供试土壤,采用大田试验,研究了半湿润农田两种杂草处理方式下(成熟后期清除杂草-A区和苗期开始清除杂草-B区),不同施氮量对夏玉米产量及氮素利用效率的影响。结果表明,当施氮量为0、45、90、1351、80 kg/hm2时,B区玉米子粒产量比A区分别增加了8.7%、12.1%、9.4%、5.0%和12.5%;吸氮量分别增加了1.5、2.9、4.85、.2和4.3 kg/hm2。A区和B区全生育期0—100 cm土层矿质氮(Nmin)累积量变化趋势基本一致,但B区比A区变幅较大。当施氮量为45、90、135和180 kg/hm2时,B区氮肥利用率、氮肥农学利用率、氮肥生理利用率均高于A区。研究还发现,在A区,当施氮量为180 kg/hm2时,杂草干生物量最大,为1518.3 kg/hm2,不施氮时,杂草的生物量最低,为845.7 kg/hm2;杂草的吸氮量随施氮量的增加而增加。可见,清除玉米农田杂草不仅可以提高作物产量和氮肥利用率,而且在减少氮素损失方面具有一定作用。

大气NH_3和介质供氮水平对不同氮效率玉米基因型叶绿素荧光参数的影响

采用开顶式气室(Open Top Chambers)进行水培试验,以两种氮效率玉米(ZeamaysL.)基因型为供试作物,通过不同大气NH3浓度处理,测定苗期各叶绿素荧光动力学参数。结果表明,供氮介质和大气NH3浓度升高对两种氮效率玉米基因型的初始荧光值(Fo)不存在显著影响。高供氮介质下,在NH3浓度升高时,氮高效5号基因型的最大荧光产量(Fm)和可变荧光(Fv)均显著减小(p<0.05),而氮低效基因型四单19的Fm、Fv值显著增加;低供氮介质下,大气NH3浓度升高对2种基因型Fm、Fv值的影响结果与高供氮介质时相反。说明大气NH3浓度升高对生长在高供氮介质下的氮高效5号基因型有一定的抑制作用,而对氮低效基因型四单19有一定程度的促进作用。在不同供氮介质下,大气NH3浓度升高时,两种氮效率玉米基因型的qN、qP值减小,说明大气NH3浓度升高时,作物对光合机构的保护能力比大气背景NH3浓度时弱。

密度对刺槐幼苗生物量及异速生长模式的影响

刺槐(Robinia pseudoacacia)具有较强的生态适应性和抗逆能力,是黄土高原植被恢复和生态重建的主要优良树种之一(程积民等,2002;陈云明等,2002)。黄土高原地区刺槐人工林密度偏大,且多为低产林(余新晓等,1996;韩芯莲等,1996;2003)。王百田等(2005)认为黄土高原刺槐单木总生物量及各部分生物量都是密度的幂函数关系,但低密度林分和高密度林分的总生物量都比中间密度林分的总生物量高。种植密度不仅影响刺槐林地生物量,而且刺槐林地出现土壤干层的部分原因就是栽植密度过大(韩芯莲等,2003),其林地生产力的维持以及可持续生长成为林草植被建设中亟待解决的问题。植物密度对种群个体数量和生长的调控可以维持种群对资源的合理利用,保持种群的稳定性(Watkinson,1980)。因生长资源的强制分配,种植密度能引起植株个体间相互作用,密度增加导致植物种内竞争产生,使种群中单株生长量和生物量发生改变(李博等,2000)。竞争的实质是植物对地上光照和地下养分资源的利用,在共享资源有限的情况下竞争会导致植物个体生长量和存活率的降低(李博等,1998)。种内竞争对植物个体生长的影响,与植物“自疏法则”的...

氮素形态和铁营养对玉米苗期生长及体内铁分布的影响

以玉米(Zea mays)品种‘豫玉-22’为材料,采用营养液培养方法,研究了低铁和正常供铁条件下供应不同形态氮素对玉米苗期生长及体内铁分布的影响。结果表明:(1)与低铁介质相比,常铁介质增加了各氮素处理玉米幼苗的株高、地上部干重、全株干重,降低了根冠比,其中硝态氮处理表现得尤其突出;与供应硝态氮(NO3--N)相比,增施铵态氮(1/2 NO3--N+1/2 NH4+-N,NH4+-N)能明显促进低铁介质中玉米生长,但在常铁介质下作用不明显。(2)相比于低铁介质,正常供铁显著提高了相应处理玉米新叶叶绿素含量及净光合速率;2种供铁介质中,NH4+-N处理的新叶叶绿素含量以及净光合速率均高于其它氮素处理。(3)相比于低铁介质,正常供铁处理总体上增加了玉米各部分活性铁含量和全铁含量,对NO3--N处理的新叶活性铁含量增加尤其明显;2种供铁介质中,NH4+-N均有利于提高新叶活性铁含量和植株地上部全铁含量。(4)玉米新叶活性铁含量不仅与其叶绿素含量显著正相关(r=0.979**),也与叶片净光合速率显著正相关(r=0.950**)。研究发现,供铁状况显著影响玉米新叶的叶绿素含量及其净光合速率且与供氮形态存在互作;供应...

燕沟流域特旱年粮食稳产示范研究

20 0 0年特旱年 ,燕沟流域通过推广抗旱保苗、调整作物种植结构、品种改良、增施肥料、大垄沟种植等技术措施 ,增产稳产效果显著 ,在全年降水量减少 36 .1%的情况下 ,粮食单产仍稳中有升 ,人均产粮达到了 5 2 5 .1kg。这对于黄土高原中部丘陵区坡改梯后保障食物安全生产、加快退耕还林还草步伐都具有一定的现实意义。

地下滴灌埋管深度对冬小麦根冠生长及水分利用效率的影响

在试验室内 ,进行了地下滴灌埋管深度对不同生育时期冬小麦根系生长和地上部分生长影响的试验研究。结果表明 :2 0 cm和 4 0 cm埋管深度的水分分布特征 ,在生育早期抑制地上部分营养生长 ,促进冬小麦蹲苗 ;而中后期则促进生殖生长 ,产量和水分利用效率较高。不考虑犁底层影响时 ,在重壤土上 4 0 cm是冬小麦进行地下滴灌的较好埋深

容器苗质量评定指标的研究

In this paper,the assessing indices of container seedlings quality were synthesized in accordance with growth and physiological factors. The results showed that the seedling quality might be determined by the dormancy state of terminal bud,nutrient condition of plant,and the state of root system. The critical quality indices should include stem diameter at soil surface,dry weight of terminal bud,and soluble sugar content. Moreover,some indices such as root system vitality and dry weight of plant were also important.

Cultivation of the intertidal brown alga Hizikia fusiformis (Harvey) Okamura: mass production of zygote-derived seedlings under commercial cultivation conditions, a case study experience

Mariculture of the brown alga Hizikia fusiformis (Harvey) Okamura as an export-oriented human food has been there more for than 20 years in China. It is now one of the five major farmed algal species along the Chinese coast. Stable and sufficient supply of young seedlings for scaling up the cultivation has been a problem throughout the farming history of this species due to the unique dioecious life cycle and relatively short time window of sexual reproduction in nature. These two factors led to a practical difficulty in obtaining zygotes at identical developmental stage in viable amounts for seedling production. A key solution to this problem is to control the synchronization of the receptacle development and to realize the simultaneous discharge of male and female gametes, such that the fertilization rate could be greatly enhanced. Focusing on one of the farmed populations in this report, we present our results on mass production of seedlings using the synchronization technique on a large scale performed in 2007. Totally 5.5 hundred million embryos were obtained from 100 kg female sporophytes. The seedlings were raised up to 3.5 mm in length in greenhouse tanks over a month and were further grown in open sea for over 3 months at two experimental sites. The success of mass production of seedlings in this alga helped to lay the basis for future trials in other species in the genus of Sargassum that have identical life cycle.

Formation and early development of tetraspores of Polysiphonia urceolata (Rhodomelaceae, Rhodophyta)

Polysiphonia urceolata is one type of potential commercial red seaweeds used for breeding and cultivation, because of its significant biochemical and biomedical application. However, the information of breeding and seedling incubation for cultivation is limited, especially the early development. In this study, tetrasporohyte and gametophyte of P. urceolata were taken as the study materials in Huiquan Bay, Qingdao, China. The cleaned and sterilized tetrasporophytes and gametophytes were pre-cultured in sterilized seawater, then nurtured at 18A degrees C, 25 mu mol photons m(-2) s(-1) in 12:12 h (light:dark) photoperiod. Continuous observation under microscope showed that the early development consists of bipolar division stage and seedling stage. In the division stage, tetraspores germinate into bipolar sporelings that further differentiate into a colorless rhizoidal portion and a lightly pigmented upright shoot. The lightly pigmented rhizoidal cell develops to a rhizoid and the larger pigmented cell transforms to an erect axis. In the seedling stage, several quasi-protuberances appear on the erect axis and form juvenile seedlings. The results demonstrate the culture of P. urceolata from tetraspores under laboratory conditions.

Temperature tolerance of young sporophytes from two populations of Laminaria japonica revealed by chlorophyll fluorescence measurements and short-term growth and survival performances in tank culture

The cold-water subtidal brown alga Laminaria japonica has been commercially fanned in the Far East and has been on top of all marine-fanned species in terms of farming area and annual output worldwide. The successful trials of transplantation of young sporophytes from the north to the south in winter along the Chinese coast in the 1950s led to the spreading of cultivation activities down to a latitude of 25-26 degrees N. Up to today, nearly 50% of the annual output of this farmed alga, as a cold-water species, comes from the sub-tropical south in China. The demand to have high-temperature-tolerant strains/ecotypes in farming area calls for a practical method to judge and select the desired parental plants for breeding programs and for seedling production. In this paper, we report our results on using chlorophyll fluorescence measurement and short-term growth performance in tank culture to estimate the temperature tolerance of offspring from two populations, Fujian Farmed Population (FFP) sampled from Fujian province (latitude: 25-26 degrees N) in subtropical area and Qingdao Wild Population (QWP) sampled from Qingdao (latitude: 36 degrees N). Contrary to what has been usually thought, the results revealed that offspring from Qingdao wild population in the north showed better performance both in short-term growth and survival rates and in optimal quantum efficiency (F-v/F-m) when exposed to higher temperature (20-25 degrees C). This result was further confirmed by fluorescence quenching analysis. QWP distributed along the southern distribution limit at a latitude of 36 degrees N in the Pacific west coast is thus taken as a more ideal one than the fanned population in subtropical region as a source of parental plants for breeding high-temperature-tolerant varieties. (c) 2006 Elsevier B.V. All rights reserved.