榄李开花生物学和繁育系统

为揭示榄李（Lumnitzera racemosa）濒危机制及开展相关保护行动，对广东珠海淇澳担杆岛省级自然保护区引种的榄李进行野外观测，观测榄李开花动态，结合运用杂交指数（OCI）、花粉-胚珠比（P/O）、花粉活力测定和柱头可授性测定、人工授粉等试验对榄李的繁育系统进行研究。结果表明（：1）榄李群体始花期为5月16日左右，盛花期为5月25日—7月1日，末花期为7月5日—8月31日，群体花期时间约106 d；花序花期持续时间为5～18 d，持续平均时间为（10.80±3.58） d；榄李单花花期持续时间为7～13 d，平均时间为（9.1±1.3） d。（2）开花当天花粉活力最高，达58.73%，第2天活力下降，第3天花粉已不具备活力；开花第1天柱头可授性最弱，后逐渐加强，第3天柱头可授性最强，达100.00%，雌雄蕊成熟时间不一致。（3）榄李杂交指数（OCI）为4，繁育系统以异交为主，部分自交亲和，需要传粉者；花粉-胚珠比（P/O）为5 740.50±1 318.20，繁育系统为专性异交；由授粉试验可知，繁育系统为自交和异交同时存在的混合交配系统。综合上述，榄李的繁育系统为以异交为主，...     

束花石斛的繁育生物学特性

为了探究束花石斛（Dendrobium chrysanthum）的繁育生物学特性,对其开花习性、花粉活力、柱头分泌特性、人工授粉结实率、自然结实种子育性和贮藏特性进行研究,结果表明：（1）束花石斛花芽萌动期为19d,现蕾期为23 d,群体花期为28 d,单花寿命10～15 d;(2)开花1～3 d花粉活力保持在90%左右,开花至第9天时花粉活力仅为23.7%,初始3 d开花的花粉活力与其他时间取样的活力差异均达极显著水平（P<0.01）,花粉活力随开花时间延长呈由强到弱的趋势;（3）束花石斛的柱头腔从开花第2天开始有分泌物,第6 d时分泌物最多,柱头过氧化氢染色结果表明在开花5～6 d时柱头可授性最强,柱头可授性随开花时间延长呈由弱到强再到弱的趋势;（4）选取花朵开放4 d的花粉和柱头人工授粉,同株异花和同花授粉未见结实。野外结实果质量2.02～4.83 g,种子长0.628～0.845 mm,宽0.082～0.110 mm,长宽比为7.798,成胚率达97.35%,种子活力为90.80%,室温条件保存6个月的种子活力维持在73.20%。上述结果说明,束花石斛单花花期不长,开花初...     

线叶忍冬开花动态特征与繁育系统研究

为了解线叶忍冬(Lonicera alberti)的开花特性及繁育系统,本研究通过野外调查,记录其花部特征、开花进程及访花昆虫特性,采用TTC、MTT、联苯胺-α萘酚和醋酸洋红四种染色方法测定花粉活力,采用联苯胺—过氧化氢法和MTT两种方法测定其柱头可授性,运用花粉/胚珠比率、杂交指数、人工授粉试验等方法检测线叶忍冬的繁育系统。结果表明:线叶忍冬花朵有花冠裂片逐一展开和花冠裂片同时展开两种开放方式;花粉在散粉后6 h活力达到最高值,柱头可授性在花朵开放后不断增强;膜翅目蜜蜂科意蜂(Apis mellifera L.)在传粉过程中起重要作用;P/O值为583.43±99.9;杂交指数为4;人工授粉套袋试验中人工异株异花授粉结实率为79.17%,自花授粉情况下结实率为9.52%。其繁育系统为混合交配系统,异花传粉需要传粉者。     

刺五加花粉活力和柱头可授性的研究

用TTC法测定了刺五加花粉的活力和寿命,用联苯胺一过氧化氢法、开花后套袋法测定了其柱头可授期,用扫描电子显微镜观察了刺五加柱头的亚显微形态特征。结果表明,刺五加长花丝植株的花粉活力比中花丝植株的高;花粉活力及其下降快慢随生境不同而有所差异;通常情况下,花粉寿命约4天。刺五加长花丝植株的柱头一直不具有可授性;中花丝植株和短花丝植株的柱头,因所处生镜不同,在开花后5-7、6-8、或7-9天具可授性;生境越郁闭,柱头具可授性的时间越晚。具可授性的柱头有独特的形态特征,即：柱头外翻、5或4裂、湿性、乳突细胞呈猫耳朵状。     

关苍术花部综合特征与繁育系统的研究

采用定点观察,运用杂交指数估算、花粉/胚珠比(P/O)测定、套袋授粉等方法,对关苍术的花部综合特征及繁育系统进行研究。结果表明:种群花期为7月下旬至9月初,单花序花期为12~17 d,小花日开放高峰期为早上7:00~8:00;花粉活力最高(90%以上)在开花后2 h以内,而此时其柱头尚无可授性,柱头在花开48~72 h以内具有较强可授性;花粉/胚珠比为6 690,杂交指数为5,繁育系统为专性异交,优势访花者为蜂类和蝇类;套袋和授粉实验结果表明关苍术不存在无融合生殖现象,自交和同株异花结实率很低,其繁育系统以异交为主。通过关苍术花部综合特征和繁育系统的研究,进一步了解其生物学特性,为关苍术资源保护和人工培育提供了理论基础。     

膏桐花粉活力与柱头可授性及其生殖特性研究

采用TTC法测定了膏桐花粉的活力和寿命,用联苯胺-过氧化氢法测定其柱头可授期,以套袋结果检测花粉活力与柱头可授性的变化,以研究柱头颜色变化与柱头可授性的关系以及开花式样对生殖成功的影响.结果表明:(1)膏桐雄花单花花期一般为2 d,花后0~9 h内花粉活力相对较高,24~33 h后花粉活力较低,48 h后花粉基本无活力.(2)膏桐雌花单花期为5~12 d,柱头可授性在花后1~4 d最强,5~8 d可授性开始逐渐降低,第9天基本失去可授性.(3)膏桐柱头表面无明显的渗出物,柱头的可授性与柱头颜色的变化相关,柱头绿色越多,柱头可授性越强;遇降雨时,少量雌花的花瓣展开而柱头仍呈球状,或者花瓣不展开而柱头先伸出,此时柱头可授性较差.(4)膏桐常表现为雄性先熟,尽管雌花数目较少,但开放速率较快,且集中在第3~5天开放,雌花开放完毕也仍有雄花未开放,为展开后的柱头留有较长时间等候昆虫传送花粉,为保障生殖成功提供了机会,但少数花序先开雌花,后开雄花,有支持异交而降低自交的倾向.     

芍药花粉活力和柱头可授性的研究

用TTC法测定了芍药花粉的活力和寿命,用联苯胺—过氧化氢法测定了其柱头的可授期。结果表明,栽培芍药单瓣花植株的花粉活力和重瓣花植株的花粉活力在开花初期(1～2d)无明显差别,但重瓣花植株的花粉活力比单瓣花植株的花粉活力下降快;通常情况下,花粉寿命约为7d。栽培品种的柱头有个体差异,单瓣花植株有的开花后2h即可分泌粘液,而重瓣花植株的柱头在开花后,有的一直不分泌粘液,有的在开花后两天才分泌粘液;单瓣花植株的柱头可授性比重瓣花植株的强,一般分别在开花后6d和3d之内具可授性。移栽的野生芍药植株的花粉活力比栽培品种下降的慢,其寿命也较长,一般为15d;柱头的可授性和粘液分泌比栽培植株强。     

八种广西特有报春苣苔属植物开花生物学特性研究

广西是报春苣苔属植物的世界分布中心,特有种较多。该研究针对线叶报春苣苔(Primulina linearifolia)、条叶报春苣苔(P. ophiopogoides)、刺齿报春苣苔(P. spinulosa)、大根报春苣苔(P. macrorhiza)、线萼报春苣苔(P. linearicalyx)、癞叶报春苣苔(P. leprosa)、桂中报春苣苔(P. guizhongensis)、百寿报春苣苔(P. baishouensis),八种广西特有种的开花物候期、开花动态、子房发育情况、花粉活性等开花生物学特性进行了研究。结果表明:(1)八种广西特有报春苣苔属植物在野外及人工环境下部分种在花期、单株开花量、单花开放持续时间具有显著性差异。人工栽培植株在单株开花量、单花开放持续时间要优于野生植株。(2)不同物种的单花花期长短不同,为3～14 d。雄蕊在花蕾阶段已经基本完成生长。雌雄蕊成熟期不一致,雄蕊先于雌蕊成熟。(3)花冠半开放至完全打开阶段的花粉活性最高,均在90%以上。(4)花冠完全开放阶段的柱头可授性最强。(5)自花授粉不能结实,异花授粉可正常结实,自交种子发芽率在80%以上。综上结果不仅为解决珍稀物种的繁殖障碍和保存种质资源提供了数据支持,而且对后续提高报春苣苔属的杂交育种成功率具有重要意义。     

七叶树花粉活力和柱头可授性变化的研究

采用花粉离体萌发法研究不同蔗糖浓度、硼酸浓度和不同温度对七叶树花粉萌发的影响及花粉活力测定,用联苯胺-过氧化氢法测定柱头可授性.结果表明:七叶树花粉萌发的最佳培养基是12％蔗糖+30 mg/L硼酸,花粉萌发的最适温度为25℃.雄花在开花当天花粉活力最高达75.69％,并在开花当天的上午10:00时,花粉活力最强,10:00～16:00花粉活力保持较高活力,是最佳授粉时段.两性花的柱头可授期持续时间较长,为8～9 d,开花3d达到最强,开花1～4 d柱头可授性保持较高水平,为授粉的最佳时间段.因此,从七叶树的花部特征、花粉活力与柱头可授性及花粉萌发的条件看,在长期的自然选择下七叶树在花部结构和开花生理上都是相配合的,以保障生殖成功.     

枸杞新品系NQ-2开花生物学研究

为探讨枸杞新品系NQ-2纯系种植结实率低的原因,采用TTC法、联苯胺—过氧化氢法、花粉—胚珠比、杂交指数和套袋试验等方法,对NQ-2花粉活力、柱头可授性及繁育系统等进行了研究。结果表明:NQ-2单花花期为3~4 d,花后8 h散粉结束,开花当天花粉活力最高,花粉寿命可持续9 d以上;开花前一天柱头已具可授性,开花第1天柱头可授性最强,开花第3天柱头基本失去可授性;枸杞新品系NQ-2平均花粉-胚珠比8615,OC I=4,结合人工授粉实验结果确定其繁育系统为专性异交。自交不亲和是导致NQ-2纯系种植落花落果结实率低的主要原因。     

酸枣生理生化特性对干旱胁迫的响应

以3年生酸枣为试材,通过盆栽控水试验法研究干旱胁迫条件下酸枣生理生化特性的响应规律.结果表明:随着干旱胁迫梯度的加大、干旱时间的延长,酸枣叶中可溶性糖、游离脯氨酸(PRO)、超氧阴离子自由基及抗坏血酸(AsA)含量累积,呈现持续上升的变化趋势;重度干旱下可溶性糖积累量高于CK 171.74％,且差异显著(P＜0.05);超氧阴离子自由基含量增加了5.77倍(P＜0.05),对细胞膜的氧化损伤加剧;PRO含量在重度干旱时最高,达到1 733.23 μg/g,是CK组的12.68倍(P＜0.01);抗坏血酸含量最高为813.10mg/100g.FW,增加了158.07％(P＜0.01).可溶性蛋白含量呈现先上升后下降的变化趋势,在中度干旱时含量最高9.99 mg/g,增加了2.11倍(P＜0.05);叶绿素a(Chl a)、叶绿素b(Chl b)及叶绿素总量呈下降的变化趋势,Chl a降低了31.08％ (P ＜0.05),Chl b降低了37.99％ (P ＜0.01),叶绿素总量降低了33.68％(P＜0.05);干旱对Chl a、Chl b的影响程度不同,导致Chl a/Chl b变化异常;类胡萝卜素含量在干旱胁迫前期上升,增加了37.72％,后期下降,降低了18.94％ (P ＜0.05).     

灵武长枣采后保鲜贮藏特性研究

以灵武长枣采后鲜果为试材,于不同贮藏条件下系统研究了其呼吸、乙烯释放、耗氧、失水及适宜贮温等特性,以期为灵武长枣贮藏保鲜措施制订提供依据。结果表明,八成熟灵武长枣在常温贮藏过程中,随贮藏时间延长,有呼吸高峰出现,乙烯释放呈双峰型;灵武长枣在果面完全着色时有呼吸升高和乙烯释放增加现象,灵武长枣可能为跃变型果实。灵武长枣采后耗氧迅速,极易失水,0℃恒温能显著延缓其后熟进程,保鲜期较常温延长21d。研究发现,控制环境低温,提高相对湿度和加强通风透气是灵武长枣贮藏保鲜的必要条件。     

枣愈伤组织诱导和再生植株

以民勤小枣幼茎和幼叶为外植体,诱导出愈伤组织,用茎尖诱导生根获得再生植株。从茎段诱导的愈伤组织属淡绿色致密的Ⅰ型和白色疏松的Ⅱ型。它们都是非胚性愈伤组织。研究了渗透压、外植体取材时间等因素对愈伤组织诱导及生长的影响。幼嫩的材料易诱导出愈伤组织,在各种基本培养基中,以３／４ＭＳ培养基诱导率最高,暗培养较光下培养有利,３％￣５％蔗糖浓度最适宜,而高渗透压不利。各种激素对继代培养的作用依次是２,４－Ｄ〉     

Chemical constituents from the fruit of Zizyphus jujuba Mill. var. spinosa

Twenty-one compounds, including ten triterpenoids (1–10), two sterols (11 and 12), six flavonoids (13–18), and three cerebrosides (19–21) were isolated from the fruit of Zizyphus jujuba Mill. var. spinosa (Bunge) Hu ex. H. F. Chou. The structures were elucidated by spectroscopic methods and by comparison of their reported spectral data. These compounds have shown the relationship between this plant and other species from the Rhamnaceae family.     

枣树体细胞胚发生和组织学研究

以临泽小枣子叶切块为外植体,在附加０．２mg/L IBA 1．0mg/L 6-BA的ＭＳ培养基上１周后切块边缘可诱导出白色胚愈伤组织,继续培养１个月后愈伤组织中产生体细胞胚。体细胞胚发生不同步,经历球形胚、心形胚、子叶胚等阶段,与合子胚发育途径相似。组织切片表面胚性愈伤组织细胞体积小,细胞核大、细胞质浓,细胞排列紧密;而非胚性愈伤组织细胞体积大、细胞核小、细胞质稀薄,子叶胚时期体细胞胚内部出现维管束,并观察到螺纹导管。     

Chemical constituents from the roots of Zizyphus jujuba Mill. var. spinosa

One new flavonoid, named (R)-2-hydroxynaringenin (1), along with fifteen known compounds, including six flavonoids (2–7), six lignans (8–13), two phenolic glycosides (14–15) and one alkaloid (16) were isolated from the roots of Zizyphus jujuba Mill. var. spinosa (Bunge) Hu ex H. F. Chou. Their structures were established on the basis of spectroscopic evidence, including 1D and 2D NMR, circular dichroism (CD) curve analysis, and by comparison with literature data. Compounds 2, 3 and 8–16 were found in Z. jujuba Mill. var. spinosa for the first time, while compounds 8, 10 and 13–15 were new to the Rhamnaceae family. In addition, the chemotaxonomic significance of the isolates was discussed in this paper.     

The Developments of Embryo and Endosperm of Zizyphus jujuba Mill

The development of the embryo of Zizyphus jujuba Mill. conforms to the Asterad type. The transverse division of the zygote results in the formation of the terminal and basal cells. The basal cell of the two-celled proembryo divides transversely first and contributes to the suspensor and a minor part of the embryo proper in further development. The terminal cell first divides vertically and the divisions in various ways contribute to almost all parts of the embryo proper. The endosperm development is of the nuclear type. Three stages may be divided as follows: the free nuclear stage, the wall formation stage and the stage in which the disorganization of the endosperm cells takes place rapidly. The developments of the embryo and endosperm are well correlated. Thus the morphological development of the embryo may be used as an indicator to tell the development stage of the endosperm. The adequate stage for endosperm culture is in the wall formation stage.     

Repetitive somatic embryogenesis and plant regeneration in Zizyphus jujuba Mill

Summary A procedure for regenerating Zizyphus jujuba Mill. (Chinese date) plants through repetitive somatic embryogenesis (RSE) was developed. Primary somatic embryos were produced from cotyledon-derived cultures of germinated plants in vitro. The highest induction frequency of primary somatic embryogenesis (PSE) was obtained with a combination of 4.52 μM 2,4-dichlorophenoxyacetic acid (2,4-D), 0.49 μM indole-3-butyric acid (IBA), and 0.44 μM benzyladenine (BA) (17.4%). These primary somatic embryos were multiplied by RSE on media with different plant growth regulator (PGR) combinations. The highest RSE frequency (51.3%), was obtained with 0.58 μM gibberellic acid (GA₃). However, the highest number (4.4 per primary somatic embryo) of repetitive somatic embryos was obtained with 0.98 μM 6-r-r-dimethylallylaminopurine (2-iP). For germination of somatic embryos, different PGRs, cold and desiccation treatment were tested. Desiccation of somatic embryos at 25±1°C for 2 wk was the best treatment for germination with epicotyl elongation and root development. Of over 256 plants regenerated, 237 (92.6%) survived.     

Agrobacterium-mediated transformation of the winter jujube (Zizyphus jujuba Mill.)

Winter jujube, a species that originated in China, is the most prominent elite variety of jujube (Zizyphus jujuba Mill.). Due to its economic value and its recalcitrance to improvements through traditional plant breeding approaches, genetic transformation techniques may have a great potential in providing the means to transfer one or more selected desirable traits into the plant genome. We reported here an improved protocol for the Agrobacterium-mediated transformation of shoot tips of winter jujube. We have identified a set of optimum transformation conditions that take into account Agrobacterium inoculum density, Agrobacterium incubation period, co-cultivation conditions, and vacuum (use of a vacuum pump to create a negative-pressure environment). The highest transformation frequency (5.2%) was obtained when the shoot-tip explants were infected for 10 min and co-cultured for 4 days with Agrobacterium at OD₆₀₀ 0.8 under a negative pressure of 0.5 × 10⁵ Pa. PCR and southern blot analyses confirmed the presence of transgenic plants and the stable integration of the target gene into the genome of regenerated plants. A histochemical staining analysis for GUS activity in the transgenic shoot tips also validated the efficiency of the transformation system.     

In vitro induction of tetraploid plants from diploid Zizyphus jujuba Mill. cv. Zhanhua

Tetraploid plants of Zizyphus jujuba Mill. cv. Zhanhua were obtained with in vitro colchicine treatment. Shoot tips from in vitro-grown plants were treated with five different concentrations of colchicine (0.01, 0.03, 0.05, 0.1, 0.3%) in liquid MS medium (Murashige and Skoog 1962), and shaken (100 rpm) at 25 °C in darkness for 24, 48, 72 or 96 h, respectively. Tetraploids were obtained at a frequency of over 3% by using 0.05% colchicine (48 h, 72 h) and 0.1% colchicine (24 h, 48 h) treatment as determined by flow cytometry. Cytological and morphological evidence confirmed the results of flow cytometric analysis. The chromosome number of diploid plants was 24 and that of tetraploid plants was 48. The stomata sizes of tetraploid plants were significantly larger than those of diploid plants, while the frequency of stomata were reduced significantly. Similarly, the chloroplast number of guard cells of tetraploid plants increased significantly. The selected tetraploid plants were grafted onto mature trees of Z. jujuba Mill. cv. Zhanhua in the field, resulted in thicker stems, rounder and succulent leaves, larger flowers and a delay in florescence time (3–4 days later) than diploid plants.