Study of Embryo and Endosperm Development of Strawberry

The fertilization and development process of embryo and endosperm of strawberry,Gelila snd Xiaoshi which were broadly cultivated in Harbin were investigated by optical microscopy, fluorescence microscopy and transmission electron microscopy.The papilla cells ranked in grid orid on its stigma and the flourishing transmitting tissue appeared in the center of its thin and long pistillary.There was only one crassinucellate hemianatropous ovule with Polygonum type embryo sac and single integument in each ovary.The germination of pollen and the pathway of pollen tube growth could be clearly observed with fluorescence microscopy:stigma→transmitting tissue in pistillary→intine of ovary placenta→the surface of ovule→micropyle.The development of embryo was consistent with most dicotyledons:zygote→2-cell proembryo→proembryo shaped like a ball→proembryo shaped like a heart→proembryo shaped like a torpedo→proembryo formed two cotyledons.The embryo was orthotropous and its development was asterad type.The endosperm development beloged to nuclear type as following:primary endosperm nuclcus→free endosperm nucleus→endosperm cell →endosperm assimilated gradually by embryo.There was a special struc-ture around it during the development of Gelila embryo.     

Analysis on Breeding Potential of 17 Maize Inbred Lines from Land Races

[Objective] The primary objective of this study was to estimate combining ability and genetic parameters of 17 new inbred lines.[Method] By using 11 commonly used maize inbred lines from Southwest China as testers and incomplete diallel cross design,combining ability and heterosis of 17 new inbred lines bred from the land races in Sichuan Basin and its surrounding areas were analyzed.In addition,the genetic parameters and heterotic groups were also investigated based on combining ability.[Result] P1-1,P1-16 and P1-14 with the better comprehensive characters can be directly used in maize breeding program.The SCA of P1-10×P2-4 was high in most traits.It is more possible to obtain high-yield hybrid combinations by the pattern Sipingtou × Lancaster.GCA variance was bigger than SCA variance in four traits such as rows per ear,grains per row,which were affected by the environments slightly and should be selected at early generations;while SCA variance was bigger than GCA variance in five traits such as 100-grain weight and plant weight,which were significantly influenced by environment and thus should be selected in later generations.[Conclusion] The paper will provide theoretical basis for further improvement of local maize varieties.     

The Inheritance and Variation of Gum Content in Guar [Cyamopsis tetragonoloba （L.） Taub.]

Guar [Cyamopsis tetragonoloba （L.） Taub.] is one of the most important industrial crops due to its richness in gum. Understanding the inheritance of gum content is a key to its successful genetic improvement. Gum content expression is reported to be controlled by additive, dominant, and epistatic effects, and modified by the environment, fg%, a relative gum content calculated by comparing with the gum content of Kinman was used as gum content （%） in this experiment. Reciprocal crosses of two lines of guar, PI 217923 and Lewis, were made to study the heritability of gum content （fg%）. fg% of four plant introductions and four commercial varieties were studied in Lubbock in 1999-2002. Estimates of broadsense heritability （h^2 b,s.） of fg% in Lewis x PI 217923 and PI 217923 x Lewis were 75.53 and 52.74%, respectively. Estimates of narrow-sense heritability （h2 n.s.） of fg% were 40.00 and 29.00% thr Lewis x PI 217923 and PI 217923 x Lewis, respectively. At least one pair of genes were estimated to control the fg% expression in these two crosses, Significant differences of fg% were found among these eight entries. PI 217923 was found to have the highest fg% among the eight entries.     

Anatomical Study on the Multi-Ovule Development and Abortion of Hanfu Apple

The terminal flower buds of 6-yr-old Hanfu apple were used to study the ovule development, ovular characteristics, cell death of abortive ovules, and dynamic change of starch grain quantity in the embryo sac with paraffin slices and terminal deoxynucleotidyl transferase-mediated fluorescein deoxyuridine triphosphate nick-end labeling （TUNEL） system. Four anatropous ovules in each ventricle could be observed before flowering. With the developing of floral organ, the bulk of normal ovules enlarged in each ventricle, the mature embryo sac differentiated into nucellus, and the egg cell developed into zygote by double fertilization. A large number of starch grains were observed during pollen tube growth and double fertilization, which guaranteed basic nutrient supply in the normal development of ovules. Moreover, abortion phenomenon of runtish ovules emerged at the stages of mature embryo sac, double fertilization and zygote development. The abortion characteristics included deformity of ovule development, degradation of nucellus tissue, separation between funiculus and ovule, abnormality of four-nucleate embryo sac, as well as development interruption of mature embryo sac. TUNEL analysis proved that ovule abortion was programmed cell death.     

Antagonizing Aspergillus parasiticus and Promoting Peanut Growth of Bacillus Isolated from Peanut Geocarposphere Soil

A set of 38 Bacillus strains isolated from peanut geocarposphere soil were screened as potential biological control agent anti-Aspergillus parasiticus. Tip-culture method with rapid and quantitative characteristics was used to determine anti-A. parasiticus activity and the process of isolation could be accelerated with this method. 12 out of 38 Bacillus strains showed high anti-alfatoxin production activity. These 12 Bacillus strains were selected to identify the characteristics of promoting peanuts germination rate. Pot experiment in greenhouse was carried out by using these strains which can promote peanuts germination. Phytohormones in the fermentation broth were also detected as another important reference factor to select the isolates as biological control agent with PGPR features. These Bacillus strains isolated from peanut geocarposphere soil not only had high ability anti-Aspergillus parasiticus, but also promoted peanut growth. Therefore, these Bacillus strains were well adapted to peanut production in the ifeld as biological control agent with plant growth promoting rhizobacteria （PGPR） features.     

Haploid Induction via In vitro Gynogenesis in Tomato （Solanum lycopersicum L.）

In order to determine the potential for haploid induction via in vitro gynogenesis in tomato, the ovules and protoplasts of embryo sacs from the hybrids Zhongza 101 and Zhongza 105 were cultured. An efficient method of ovule isolation was established in this study. Using this method, 100-150 ovules could be isolated from one ovary. Isolated ovules were cultured on three induction media to induce gynogenesis in vitro. During culture, ovules were enlarged markedly, with opaque white color. When observed microscopically, there were cell divisions and cell clumps in embryo sacs. Subsequently, the cell clumps in embryo sacs ceased growth, likely because the integument grew faster than embryo sacs did and hindered the fiarther development of embryo sacs. Therefore, subsequent callus morphogenesis might be originated from the integument. Thousands ofcalli from the two tomato varieties were obtained. Five diploid plants were regenerated after 15 months of subculturing. To eliminate the hindering effect of integument on embryo sac cells, the protoplasts of embryo sacs were prepared and cultured. After 48 hours of culture, the protoplasts of embryo sacs doubled in size and gradually formed clusters of cells. These results suggested that gynogenesis might be a potential way for haploid induction in tomato.     

茄类内生菌的分离及拮抗细菌的筛选(英文)

[Objective] The aim of this study was to isolate the endophyte of three solanaceae fruits and vegetables such as tomato, pepper and eggplant, to screen and identify the bacterial wilt antagonistic bacteria. [Method] According to the lapping liquid culture method, the endophyte of three plants was isolated by the selective medium and purified by the plate streaking method, so the purified endophyte was screened by the hyphal pieces confront culture method. Furthermore, the screened antagonistic and endophyteic bacteria was identified and classified through culture characteristics of isolates and morphological features of thallus, Gram stain as well as physiological and biochemical reactions. [Result] Fifty-three endophytic bacteria, fifty-three endophytic fungi and forty-four endophytic actinomycetes were separated from the endophyte of three plants. The screened fourteen endophytic bacteria with strong antagonistic effect on the bacterial wilt were classified to Bacillus, Escherichia, Klebsiella, Agromonas, Erwinia and Curto Bacterium respectively. Especially, Bacillus was the dominant species, which had the strongest antagonistic effect on the bacterial wilt. [Conclusion] This study provides an effective way for biological control of the bacterial wilt in solanaceae.     

水牛子宫内膜腺上皮细胞和基质细胞的分离·培养和生物学特性研究(英文)

[Objective] The experiment aimed to set up a method for isolating and culturing endometrial stromal cells (BESC) and endometrial glandular epithelial cells(BEGEC) of buffalo as well as laid foundation for studying biological mechanism of embryo implantation and uterine diseases. [Method] The enzymatic digestion method, scraping method, serial filtration and differential velocity adherent technique were used to isolate BESC and BEGEC, then immunocytochemical method and TRYPAN-Blue assay were used to determine the purity and survival rate of isolated cells. [Result] The BESC and BEGEC were successfully isolated and cultured while immunocytochemical method and cell count method demonstrated that the purity was over 90%. The result of TRYPAN-Blue assay shown that survival rate of BESC and BEGEC was 91% and 78% respectively. [Conclusion] The enzymatic digestion method, scraping method, serial filtration and differential velocity adherent technique could isolate BESC and BEGEC with high purity.     

拮抗链霉菌AFLP分析技术体系的研究(英文)

[Objective] The aim of this study was to investigate the preparation method and amplification system of antagonistic streptomyces DNA templates based on AFLP assays, and also provide a basis for the application of AFLP technology in the analysis of streptomyces or even actinomyces. [Method] The DNAs were extracted by the modified CTAB method and amplified by the Pst Ⅰ/Mse Ⅰ AFLP kit and its reaction system. The amplified products were analyzed by the denatured polyacrylamide gel electrophoresis. [Result] The genomic DNAs of ten antagonistic strains of Streptomyces were extracted and tested. The result of 0.8% agarose gel electrophoresis showed that the major DNA bands were clear without degradation and RNA residue, with the fragment sizes ranging from 37.64 to 40.86 Kb. By ultraviolet spectrophotometry, the OD260/OD280 values varying from 1.625 to 1.833 were obtained. Furthermore, the agarose gel electrophoresis of DNA products digested by Pst Ⅰ/Mse Ⅰ presented the dispersed fluorescent long band, which indicated that the enzymatic hydrolysis was fully carried out. The amplified bands of DNA templates by the screened three pairs of primers were clear with rich polymorphism. [Conclusion] The preparation method and amplification system of DNA template established in this study can be used in the AFLP analysis of Streptomyces.     

棉花黄萎病拮抗细菌7-30菌株的筛选与鉴定(英文)

[Objective]The aim of this study was to screen the antagonistic spore-forming bacteria of Verticillium dahliae and identify its physiological and biochemical characteristics.[Method]Taking the cotton verticillium wilt pathogen Verticillium dahliae V-190 as the test microorganism,the antagonistic spore-forming bacteria were screened.Through the preliminary screening and secondary screening,an antagonistic bacteria strain 7-30 with stronger antibacterial activity was obtained,and its morphological characteristics,physiological and biochemical characteristics were also identified.[Result]84 antagonistic bacteria strains were isolated from soil in various places by the preliminary screening.Especially,18 strains with better antagonistic ability were screened again,so an antagonistic bacteria strain 7-30 with the diameter of inhibition zone 18.9 mm and stronger antibacterial activity was obtained.According to its morphological characteristics,physiological and biochemical characteristics,the antagonistic bacteria strain 7-30 was identified as Bacillus subtilis primarily.[Conclusion]The strain 7-30 was obtained as the antagonistic spore-forming bacteria of Verticillium dahliae.     

一种新的非酶法分离荠幼胚的方法

1材料与方法1．1材料试验所用荠的花和幼嫩果实，于2009年2月下旬至3月上旬采自河南省洛阳市。采集后，在花序中、上部选取花已开败的幼嫩果实进行解剖，或将其浸泡于60％的乙醇溶液中备用。解剖前取出，再将其浸泡于蒸馏水（或自来水）中除去乙醇，以减缓花、果在解剖过程中的干缩进程。     

一种新的非酶法分离荠幼胚的方法

[目的]用非酶法手工分离荠的胚珠和幼胚等微小结构。[方法]应用胶块和解剖镜等普通工具,手工、非酶法分离并观察荠的花形态及荠胚珠和幼胚。[结果]荠花具有4片花萼、4片花瓣、四强雄蕊,短角果;荠的胚发育过程包括球形胚、心形胚和鱼雷形胚3个发育阶段。[结论]利用胶块的黏性,能使植物胚珠和胚等微小结构的手工分离操作变得比较简单,具有对幼胚损伤小、能保持原有形态等优点。可用于多领域的研究。该类工作在实验设施较差的实验室也能完成。     

无核葡萄胚挽救技术研究

[目的]研究无核葡萄胚挽救过程中的一些主要影响因素,为今后开展无核葡萄胚挽救育种提供参考。[方法]以6个无核葡萄杂交及自交组合未成熟胚珠为试材,研究了接种时期、培养基及激素浓度对无核葡萄胚挽救效果的影响。[结果]各组合适宜的胚珠接种时期为：火星无核×无核白鸡心在花后40d,金星无核×康能无核在花后55d,优无核×布朗无核在花后45d;采用ER为基本培养基更有利于胚珠的发育和萌发;不同品种的胚珠对培养基中IBA浓度要求不同,幼胚发育程度较高,败育时期较晚的胚珠,适合较低的IBA浓度,胚发育程度较低,败育较早的胚珠则需要较高的IBA浓度。[结论]接种时期、培养基种类和培养条件对无核葡萄胚挽救的成功都非常重要,且各条件的确立要视具体品种而定。     

3, 5-二硝基水杨酸比色法测定荠菜多糖含量的研究

[目的]为荠菜多糖的开发提供一定的理论依据。[方法]采用3,5-二硝基水杨酸（DNS）比色法测定了荠菜还原糖的含量并通过单因素试验研究确定了其最佳测定条件。[结果]在0．5～1．5ml的范围内,吸光度随着DNS用量的增加而增大;当DNS的用量大于1．5m1时,吸光度基本不变。在4～6min内,吸光度随着加热时间的延长而显著增大;6min后吸光度基本稳定。在70～90℃内,吸光度随着反应温度的升高而显著增大;反应温度超过90℃后,吸光度随着反应温度的升高而趋于稳定。放置20min后吸光度达到最大。3,5-二硝基水杨酸比色法测定的荠菜还原糖含量为1．002％,总糖含量为7．3393％,多糖含量为6．3373％。[结论]该试验确定的DNS比色法的最佳测定波长为510nm,最佳DNS用量为1．5ml,最佳反应温度为90℃,最佳反应时间为6min,最佳放置时间为20min。     

反季节杧果花器官结构对胚胎败育的影响

[目的]研究反季节杧果花器官结构对胚胎败育的影响。[方法]观察反季节杧果生产中花器官和胚胎的结构,探索反季节杧果胚胎败育过程中"金煌"品种的细胞学结构变化,研究胚胎败育的细胞学机理。[结果]"金煌"雄蕊退化,花粉囊间发育差异十分显著,花粉囊小的花粉数量少,而花粉囊大的花粉数量多且发育至成熟,仅有少部分花粉囊的花粉退化,花粉生活力为76.9%。雌器官出现子房萎缩变形、胚珠横生现象;胚珠周围的输导组织断裂现象普遍;在胚囊8核期间,在同一或不同胚珠中出现极核细胞消失或者退化,反足细胞个别退化、助细胞个别或者全部退化的现象。[结论]以上因素都会影响胚珠受精,引起授粉受精不良。     

融安滑皮金柑种胚发育的细胞学观察

【目的】观察不同发育时期融安滑皮金柑种胚发育情况,探讨其败育的原因,为该品种资源的创新利用及金柑育种研究提供科学依据。【方法】以融安滑皮金柑为试材,采用石蜡切片、植物显微技术等方法,对其胚珠发育过程进行系统的观察研究,并对成熟果实种子败育情况进行统计分析。【结果】融安滑皮金柑在授粉阶段和胚囊发育阶段表现正常;胚珠发育至花后7d,一部分因极核未能正常受精而导致胚胎败育,另一部分因内、外珠被发育时期紊乱使珠心和胚囊发育受到扰乱而导致胚胎败育。融安滑皮金柑成熟果实中,每果种子数5～9粒,其中饱满种子（2．3±0．17）粒,败育种子（9．7±0．17）粒,败育率（80．8±1．45）％,正常种子为多胚性。【结论】融安滑皮金柑种子败育属于中途败育类型。     

蓝猪耳大孢子发生和雌配子体发育

[目的]研究蓝猪耳大孢子发生和雌配子体发育。[方法]采用常规石蜡切片和超薄切片技术研究蓝猪耳大孢子发生和雌配子体发育过程。[结果]蓝猪耳为中轴胎座,倒生胚珠,单珠被,薄珠心。大孢子母细胞减数分裂形成T-形和直线形四分体。T-形四分体合点端大孢子具功能,胚囊为蓼型发育形式;直线形四分体中珠孔端大孢子具功能,最终形成八核七细胞的成熟胚囊。胚囊发育异常偶有发生,但大部分胚囊都能发育成正常的7-细胞胚囊。[结论]蓝猪耳雌性生殖细胞发育异常并不是其结实率低的主要原因。     

无核葡萄胚败育时期的确定及接种时间对胚萌发的影响

研究了4个无核葡萄品种果实发育过程中,果实及胚珠质量的动态变化情况以及百乐无核葡萄不同接种时期对胚发育和成苗的影响.结果表明:早熟品种火焰无核和1124,中熟品种百乐无核以及晚熟品种鲁贝无核的胚败育时期分别在胚发育35、404、5 d后;且百乐无核在胚发育40 d接种的胚珠发育率(64.7%)和成苗量(18株)最高.     

牛角瓜大孢子与小孢子的发生发育观察

【目的】明确牛角瓜[Calotropisgigantea(Linn.)W. T. Aition]大孢子与小孢子的发生发育情况,为其耕作栽培学及植物学研究提供基础资料,也为其遗传育种研究及品种改良打下基础。【方法】于盛花期采集不同发育时期的牛角瓜花蕾,运用石蜡切片技术观察其大孢子与小孢子的发生发育特征。【结果】牛角瓜大孢子与小孢子的发育时期与其花器官外部形态特征存在对应关系,雄蕊的发生发育早于雌蕊。牛角瓜小孢子黏连成块状结构,与普通植物的分散型小孢子形成鲜明对比,其小孢子母细胞发育为连续型,绒毡层细胞有3~4层,为腺质绒毡层,单核期绒毡层细胞壁开始降解,花粉成熟后绒毡层细胞自溶。胚囊发育类型为蓼型,胚珠横生,无珠被;四分体呈线型排列,成熟胚囊有次生核。【结论】牛角瓜大、小孢子发生及雌、雄配子体发育与花器官外部形态大小密切相关,其结实率低并非雌性或雄性败育所致。     

枣幼胚培养中褐化的研究

研究了建立枣幼胚离体培养体系时品种、外植体、活性炭、接种方式和基本培养基对褐化的影响,其中,外植体是枣幼胚离体培养时褐化的关键因素。取出早脆王胚珠,剥去珠被接种可以有效防止褐化。不同发育时期的胚珠褐化不同,其结果与品种及接种方法有关。完整接种的早脆王胚珠随其成熟度的增加褐化加重,但切开远胚端接种金丝丰和冬枣的幼嫩胚珠更容易褐化。加倍的MS培养基可以促进早脆王幼胚褐化,附加活性炭可以减轻早脆王完整胚珠褐化。