Male Sterile Lines of Zinnia elegans and Their Cytological Observations

In order to find out a new pathway for utilizing heterosis of Zinnia elegans and accelerate breeding process, the mechanism of anther development of a male sterile line was explored. Backcross, sibmating, selling of fertile plants and testcross with inbred lines were analyzed and identified in the field, and cytology was observed. Recessive nucleus male sterile line AH209AB capable of being a maintainer was obtained by successive backcrosses with male sterile plants and fertile F1 plants as male parents. Cytological and anatomical studies indicated that： （1） The wall of normal anther was constituted of four layers of cells such as epidermis, powder chamber wall, middle level and tapetum cells. The process in meiosis of pollen mother cell in Zinnia elegans was normal and cytoplasm divided simultanously. Mature pollen grain was tricellular type. （2） The petal of male sterile plant degraded as a thread-like structure, the stamens were villiform in appearance and no pollens were formed. The result showed that the anther of male sterile plant no longer proceed to differentiate spore mother cell and the pollen sac after the formation of the tissue of sporogenous cells, there was no evident boundary between tapetum cell, middle lamella and inner wall of PMC, tapetal cells did not develop from the very beginning. So the abortion type was completely structural male sterility. The male sterile line belongs to non-sporange male sterile type and is of great use in F1 seeds production.

Construction and Analysis of Suppression Subtractive cDNA Library from Fertile Disk Floret Buds of Zinnia elegans

In order to isolate genes related to anther development and understand molecular basis of male sterility,cDNA library of Zinnia elegans was constructed using suppression subtractive hybridization（SSH） approach.672 different expressed clones were selected from the fertile disk floret buds.PCR results showed that cDNA inserts were ranged from 100 to 750 bp.303 positive clones screened by dot-blot hybridization were sequenced.273 out of 303 sequenced clones produced readable sequences;these sequences represent 87 non-repetitive sequences.The homology alignment showed that 76 expressed sequence tags（ESTs） had functional annotations in GenBank,the other 11 ESTs without any homology to the known gene.In addition,87 ESTs were divided into 17 groups according to MIPS of Arabidopsis thaliana database.Sequence data from the cDNA library have been deposited with the GenBank under the accession numbers GT067016-GT067085.As an important result in this study,7 genes related to anther development were isolated.Results from semi-quantitative RT-PCR showed 6 genes were expressed only in disk florets of fertile plants compared with that of male sterile plants.These ESTs obtained provide important clues for further isolation and identification of fertility-related genes in Z.elegans.

Effects of 5-Aminolevulinic Acid (ALA) on Zinnia hybrida Growth and Phytoremediation Effects in Oil-Contaminated Soil

In this study, we compared plant height, weight, soil TPH concentration, and soil DHA level after 18 weeks of Zennia hybrida cultivation with four different concentrations of 5-Aminolevulinic acid (ALA)-based liquid fertilizer: 1500-fold, 5000-fold, and 8000-fold dilutions, along with a non-treatment control of diluted ALA. The plants of ALA-treated were significantly taller than the non-treatment control. The plants of ALA-treated plants were higher in shoot fresh weight, shoot dry weight, and root dry weight than the non-treatment control. The plot of ALA-based liquid fertilizer with the 5000-fold dilution was significantly highest in shoot fresh weight, shoot dry weight, and root dry weight. ALA-treated plants were lower in the soil Total Petroleum Hydrocarbon (TPH) concentration than the non-treatment control. The plot of ALA-based liquid fertilizer with the 5000-fold was significantly lowest in the TPH concentration. In addition, ALA-treated plants were higher in the soil dehydrogenase activity (DHA) than the non-treatment control. The plot of ALA-based liquid fertilizer with the 5000-fold was significantly highest in the TPH concentration. This study indicated that ALA-applied zinnia-grown oil-contaminated soil is more effective than not. The remediation in oil-contaminated soil with ALA-based liquid fertilizer is more effective than the non-treatment control;furthermore, ALA application with 5000-fold dilution was most suitable in oil-contaminated soil among other plots.

Application Effect of Sludge Extract in the Planting of Zinnia elegans

[Objectives] The research aimed to study the effect of sludge extract on the growth of Zinnia elegans, and evaluate its possibility of application. [Methods] Z. elegans was taken as research object, and the effect of sludge extract on the growth indicators of Z. elegans was studied. [Results] The sludge extract can obviously promote the growth of Z. elegans, and the growth indicators were significantly improved when compared with the control group. The application effect was close to that of foliar fertilizer purchased in the market. In each treatment group, the low concentration of sludge extract did not significantly promote the growth of Z. elegans, while too high concentration could inhibit its growth, and the optimal dilution ratio was 600 times. [Conclusions] Further preparation of amino acid foliar fertilizer from sludge extract and its application in planting garden plants can realize the resource utilization of sludge and has a good development and application prospect.

Downregulation of the Basic Peroxidase Isoenzyme from Zinnia elegans by Gibberellic Acid

Hypocotyl formation during the epigeal germination of seedlings is under strict hormonal regulation. In a 3 d old Zinnia elegans seedling system, gibberellic acid （GA3） exerts an opposite effect to that exerted by light on hypocotyl photomorphogenesis because GAz promotes an etiolated-like growth with an inhibition of radial （secondary） growth. For this reason, the effect of GA3 on the basic peroxidase isoenzyme from Z. elegans （ZePrx）, an enzyme involved in hypocotyl lignin biosynthesis, was studied. The results showed that GA3 reduces ZePrx activity, similarly to the way in which it reduces seedling secondary growth. This hormonal response is supported by the analysis of the ZePrx promoter, which contains four types of GA3-responsive cis.elements： the W Box/O2S; the Pyr Box; the GARE; and the Amy Box. Taken together, these results suggest that ZePrx is directly regulated by GA3, with this effect matching the inhibitory effect of GA on the hypocotyl secondary growth.

百日草的核形态学研究

研究百日草(Zinnia elegans Jacq.)的核形态学特征,可为分类和育种工作提供细胞学依据。采用常规压片法,对百日草进行了核形态学研究。结果表明,1间期核里染色中心数目较少,形状比较规则,多分布于细胞中央,为复杂染色中心型。2有丝分裂前期染色体由染色较深的异固缩节段和染色较浅的常染色质节段相间排列而成,较多地分布于细胞中央,前期染色体为中间型。3有丝分裂中期染色体数目2n=24,为二倍体。4核型公式为:K(2n)=2x=24=24 m,最长染色体与最短染色体的比值为1.52,臂比大于2的染色体百分率为0,核型属1A型,为最对称的核型。臂指数NF值为48。5有丝分裂后期和末期正常。这些核形态学的证据进一步支持了百日草在形态上属于原始类群的观点.

百日草柱头可授性和花粉生活力的研究

【目的】观察百日草花后柱头的形态变化,测试百日草的柱头和花粉的寿命,为提高百日草的人工授粉效率和杂交育种提供依据。【方法】采用联苯胺-过氧化氢测定柱头可授性、花粉离体萌发试验和田间试验相结合的方法进行研究。【结果】(1)百日草在8:30～16:00开花,花粉生活力为18h;(2)舌状花柱头保持膨胀,花柱挺立可达7～10d;(3)联苯胺-过氧化氢测定柱头可授性的结果显示,花后1～3d可授率为79.9%～83.1%,4～6d为56.1%～63.0%,7～10d为37.0～43.3%;(4)丫状时期、丫状尖端稍弯曲时期、柱状时期田间授粉的结实率分别为76.3%、48.5%和36.8%;(5)未经处理的柱头结实率为48.8%,去丫状一裂者为30.9%,去柱头只留花柱者为19.9%。【结论】(1)用联苯胺-过氧化氢方法测定百日草柱头可授性可行;(2)花后2～3d柱头丫状时期进行人工授粉结实率高;(3)柱头顶端的毛状突起增大了吸收花粉的表面积,有利于授粉结实。

水杨酸对盐胁迫下百日草种子萌发及幼苗生理特性的影响

以百日草品种“芳菲1号”为试材,在100mmol/L NaCl胁迫处理下,研究了不同浓度（0,0.5,1.0,1.5和2.0mmol/L）水杨酸浸种对百日草种子萌发以及不同浓度水杨酸（SA）注根和叶面喷施对幼苗生长及生理特性的影响。结果显示,SA一定程度上可以缓解盐胁迫对百日草种子萌发和幼苗生长的影响。其中,0.5,1.0,1.5mmol/L SA处理的种子具有较高的发芽势、发芽率、发芽指数、活力指数;幼苗株高、茎粗、根冠比增加;叶片叶绿素含量明显升高;超氧化物歧化酶（SOD）、过氧化物酶（POD）活性以及脯氨酸、可溶性糖、可溶性蛋白含量均变高,丙二醛（MDA）含量降低。表明0.5～1.5mmol/L的SA处理通过促进百日草叶片叶绿素合成,保护其光合作用,维持了植物的正常生长;同时,通过调节百日草抗氧化及抗渗透胁迫能力,有效缓解了盐毒害,尤其以1.0mmol/L效果最佳,2.0mmol/L的SA处理未见有明显缓解。

外源水杨酸对百日草生长及开花的影响

为探讨外源水杨酸(SA)对百日草(Zinnia elegans Jacq.)生长发育的影响,以梦境百日草为试验材料,研究了不同浓度SA对百日草生长及开花的影响.结果表明,0.5mmol/L水杨酸处理显著提高百日草的株高、生物量、真叶数及开花数量,1.0mmol/L处理与对照相比大多差异不显著.综合来看,适宜浓度的外源SA对百日草的生长及开花有明显的促进作用.

细叶百日草对镉的生长响应及富集特征研究

为探讨观赏绿化植物应用于植物修复技术的可行性,以细叶百日草(Zinnia angustifolia(Z.linearis))为试验材料,通过盆栽模拟试验,研究其在不同浓度镉处理下的生长响应和富集特征。结果表明,镉处理浓度≤60 mg·kg-1时对细叶百日草的生长有一定的促进作用,表现为植株生物量的显著增加,当镉处理浓度达到120 mg·kg-1时,植株生物量有所降低,但是与对照无显著差异。细叶百日草各器官中镉含量表现为根>茎>叶>花的分布格局,且随着镉处理浓度的增加均逐渐增加,在镉处理浓度为120 mg·kg-1时达到最大值,分别为462.47、271.86、210.36、116.38 mg·kg-1。植株对镉的富集系数均大于1,地上部对镉的最大富集量每盆为1 342.44μg·pot-1,每株为335.61μg·plant-1。说明细叶百日草对镉有很强的耐性和富集能力,是一种良好的修复镉污染土壤的观赏绿化植物。本文为观赏绿化植物应用于重金属污染土壤修复提供一定依据。

水分胁迫下烯效唑对百日草幼苗光合特性及叶解剖结构的影响

以百日草‘芳菲1号’为试材,研究不同水分胁迫下烯效唑(S3307)对其幼苗生长、光合特性及叶解剖结构的影响,以明确S3307对百日草的抗旱作用及其机理。结果显示:(1)在水分胁迫下,百日草的生长均受到不同程度的抑制,叶绿素含量显著降低,光合作用受到抑制,叶解剖结构有所变化。(2)S3307处理后,均能够显著降低所对应的不同程度水分胁迫下百日草的株高,显著增加茎粗、叶面积、叶片厚度、栅栏组织厚度和根冠比,显著增加叶绿素含量,提高百日草的净光合速率(Pn)、气孔导度(Gs)、蒸腾速率(Tr)和水分利用效率(WUE)。研究表明,S3307能够提高百日草的抗旱性,而且在轻度和中度水分胁迫下Pn的下降主要是由气孔因素引起,而在重度水分胁迫下光合速率的下降是由非气孔因素引起的。

百日草炭疽病病原菌的分离鉴定及防治药剂筛选

为明确百日草炭疽病的病原菌,依据柯赫氏法则,对采集的病叶进行病原物分离并进行分离菌株致病性试验。确定菌株的致病性后,结合菌落形态,初步判断其为炭疽菌。运用分子生物学方法,扩增获得肌动蛋白基因(actin gene, ACT)、几丁质合酶基因(chitin synthase A gene, CHS)、磷酸甘油醛脱氢酶基因(glyceraldehyde-3-phosphate dehydrogenase, GAPDH)、核糖体内部转录间隔区(ribosomal internal transcribed spacer, ITS)、锰超氧化物歧化酶(manganesesuperoxide dismutase, SOD2)、谷氨酰胺合酶(glutamine synthatase, GS)、微管蛋白(beta-tubulin-TUB2)以及钙调蛋白(calmodulin, CaM) 8个基因序列,进行联合系统发育分析,确定该菌为暹罗刺盘孢菌Colletotrichum siamense。离体抑菌活性测定结果表明,9种杀菌剂制剂75%肟菌·戊唑醇水分散粒剂、35%氟菌·戊唑醇悬浮剂、75%百菌清可湿性粉剂、250 g/L嘧菌酯悬浮剂、50%咪鲜胺可湿性粉剂、10%苯醚甲环唑水分散粒剂、10%丙硫菌唑悬浮剂、500 g/L氟啶胺悬浮剂和25%溴菌晴可湿性粉剂对供试病原菌的EC50值分别为0.152、0.407、2.48、252、0.034 2、0.556、317、0.002 91和27.4 mg/L,其中500 g/L氟啶胺悬浮剂和50%咪鲜胺可湿性粉剂抑菌活性最强。

百日草雄性不育系的获得及其细胞学观察

【目的】获得百日草雄性不育系,并探索其花药发育机制,为百日草杂种优势的利用提供一条新途径。【方法】采用自交F2代出现的不育材料连续回交获得的不育系再进行兄妹交、可育株自交,结合与多个自交系测交进行不育类型鉴定,并对其细胞学进行研究。【结果】获得了百日草深红色单隐性雄性不育两用系AH209AB。正常可育的花药,其药壁由4层细胞构成,即表皮、药室内壁、中层和绒毡层;花粉母细胞减数分裂过程中的胞质分裂为同时型;成熟花粉为三细胞型。雄性不育株花瓣退化或丝状,雄蕊退化成丝状,内无花粉,外观呈绒毛状;花药在造孢细胞组织形成后,没有继续分化产生药壁结构、花粉母细胞及其花粉,为完全的结构性雄性不育类型。【结论】该雄性不育系属于无花粉囊型不育系,在F1代种子生产上具有重要的利用价值。

高温胁迫下不同浓度烯效唑对百日草幼苗光合作用和叶绿素荧光参数的影响

以百日草3~4叶期的穴盘苗为试验材料,分析了高温胁迫下不同浓度（10,20,30,40 mg/L）烯效唑（S3307）对百日草幼苗的同化产物分配、光合作用、水分利用率、叶绿素荧光参数等指标的影响。结果表明：高温胁迫下S3307处理的百日草幼苗的株高降低,茎粗增加,地上部冠干重、地下部根干重及其根冠比均大于对照,并且S3307处理还提高了百日草幼苗的净光合速率、叶片水分利用率,显著减缓了高温胁迫对百日草幼苗光合系统Ⅱ（PSⅡ）的抑制作用,使百日草幼苗PSⅡ最大光化学效率（Fv/Fm）、实际光化学效率（ΦPSⅡ）、光化学猝灭系数（qP）下降缓慢,非光化学猝灭系数（NPQ）升高幅度增大,从而使光合机构免受高温胁迫的伤害。以30 mg/L的S3307处理效果最佳。

植物生长调节剂对百日草生长及开花的影响

以百日草(Zinnia elegans Jacq.)为试材,采用不同浓度的赤霉素(GA3)、油菜素内酯(BR)和不同稀释倍数的的花粉酵素对百日草进行2次全株喷施处理,探讨对百日草花期及开花质量的影响。结果表明,在一定范围内喷施赤霉素可使百日草开花提前、延长花期,而喷施油菜素内酯、花粉酵素可不同程度地推迟百日草开花;三者均可促进花径、叶面积、株高、单株花朵数的增加,提高百日草的观赏价值。综合考虑,3种植物生长调节剂中以0.02 mg/L的油菜素内酯喷施处理的效果最好;其次是花粉酵素800倍稀释液处理;赤霉素喷施浓度以100 mg/L为宜。

百日草自交系花粉萌发条件及花粉活力研究

研究了不同培养基、采集时间、贮藏温度和湿度等因素对百日草花粉离体培养萌发效果的影响。结果表明:花粉萌发较适宜的培养基为ME3+16%PEG4000+12%蔗糖;上午10:00采集花粉萌发率要显著地高于下午3:00采集的;在4℃湿润条件下贮藏18 h,J6花粉生活力下降较小,而在室温干燥条件下几乎失去活力;相对湿度在66%以下时极不利于花粉保存;11个百日草自交系花粉离体萌发率差异显著。

大批量生产百日草链格孢菌孢子的技术

通过研究光照、菌丝体粉碎、培养基等因素对百日草链格孢菌孢子形成的影响，试图探寻大批量生产该菌孢子的条件和方法。结果显示，低氮培养、近紫外光照射、适当适时的菌丝体粉碎能诱导百日草链格孢菌形成大量的孢子。在上述条件下，应用液体发酵培养菌丝体，结合固体培养诱导孢子，是一种可行的方法。

百日草根尖和花药愈伤组织染色体制片技术

以种子萌发根尖和花药愈伤组织为材料,研究了取样时间、预处理方法对百日草染色体制片的影响。结果表明:根尖上午8:00～9:00,花药愈伤继代3～5d上午9:00～10:00为最佳取样预处理时间;采用三种药剂预处理活体根尖,以4℃下饱和对二氯苯溶液或0.002mol/L的8-羟基喹啉液预处理8h效果最佳,花药愈伤则以饱和对二氯苯溶液预处理6h效果最佳。本实验的预处理温度是固定的,可克服预处理随季节和时间温度的变化而带来的不稳定性,且百日草花药愈伤染色体观察为首次报道。

百日草链格孢菌毒素对加拿大一枝黄花叶片伤害的生理生化研究

采用植物抗性生理和光合生理的分析测定技术,研究了分离自罹病苍耳植株上的百日草链格孢菌产生的毒素对加拿大一枝黄花叶组织细胞膜透性、丙二醛(M DA)含量、光合能力以及过氧化物酶(POD)、抗坏血酸过氧化物酶(APX)和过氧化氢酶(CAT)活性的影响.结果表明,百日草链格孢菌毒素使加拿大一枝黄花叶组织细胞膜透性上升,膜脂过氧化加强,M DA含量上升;叶片的POD、APX和CAT的活性均较对照降低,其光合作用明显地受到抑制.说明百日草链格孢菌毒素具有防除加拿大一枝黄花的潜力.

百日草链格孢菌粗毒素的生产、提取及稳定性的研究

百日草链格孢菌是从世界性恶性杂草苍耳罹病植株上分离得到的植物病原真菌，研究表明该菌对紫茎泽兰有致病作用，可产生致病的次生代谢产物-毒素。对该菌的次生代谢产毒能力情况进行了研究。对培养基条件进行了筛选，确定了有利于该菌产毒的温度为25℃、pH值为6．5、培养时间为9-12d、转速110r／min、黑暗与充分溶氧等培养条件。利用大规模发酵与发酵产物后提取工艺获取粗毒素，并通过分析贮存时间、温度、光照对粗毒素稳定性的影响，表明其具有开发为天然微生物源除草剂的潜力。