两个新发现的黄瓜性别决定基因遗传规律的研究

 选取黄瓜两个雌雄同株型的强雌性高代自交系材料与普通雌雄同株、两性花株、纯雌株高代自交系材料杂交, 通过对F₁、F₂、BC₁ -1 和BC₁ -2 代性型观察, 统计分析表明: 雌雄同株型的强雌性由1对不完全显性和1对隐性基因控制, 暂定名为Mod-F₁和mod-F₂, 它们是新发现的黄瓜性别表达的两个基因, 能增强黄瓜植株雌性的表达, 与F和M 基因独立遗传。     

Ethylene evolution during germination of different sex phenotypes of cucumber

Ethylene production by germinating seeds of cucumber during the first 4 days after imbibition was determined for different sex phenotypes. Ethylene emanation from germinating seeds of gynoecious plants was much higher than from those of the more male phenotypes of androecious, monoecious and hermaphrodite. Ethylene addition and removal (by hypobaric ventilation) during germination caused no apparent changes in subsequent sex expression of the plants. Ethylene production by single F₂ seeds was not closely associated with subsequent sex expression of the plants derived from a cross between gynoecious and androecious cucumbers. These studies suggested that ethylene production during germination was not always closely related to subsequent sex expression.     

不同基因型黄瓜性别与乙烯释放速率的关系

采用气相色谱法测定全雄株(ffMMaa)、雌雄同株(ffMMAA)、全雌株(FFMMaa、FFMMAA)和两性花株(FFmmAA)不同基因型近等基因系材料间的乙烯释放速率,探讨乙烯与黄瓜性型的关系。结果表明,不同近等基因系乙烯释放速率之间存在显著性差异。全雄株78♂(ffMMaa)的乙烯释放速率较低;雌雄同株406(ffMMAA)四叶一心期的乙烯释放速率大于两叶一心期;含6.33个F基因拷贝数的雌性系10098♀(FFMMaa)与F基因拷贝数为2.15的雌性系WI1983G(FFMMAA)相比,乙烯释放速率较低;雌性系WI1983G(FFMMAA)的乙烯释放速率极显著高于两性花株WI1983H(FFmmAA),两性花株WI1983H又极显著高于雌雄同株WI1983GM(ffMMAA);说明黄瓜性别表型与内源乙烯密切相关。从试验结果推断:雄株(ffMMaa)中乙烯释放速率低可能与A基因失活相关,雌性系(FFMMaa)乙烯释放速率可能受A/a基因的调控,两性花株(FFmmAA)中虽然M基因失活但内源乙烯释放速率并未降低很多,可能F基因产生的内源乙烯没有直接作用到两性花的雄蕊上。     

甜瓜不同性别花蕾发育中激素和多胺的变化

 以甜瓜雌雄异花同株和雄全同株近等基因系为材料，采用高效液相色谱(HPLC)技术，分析雌花、雄花和两性花4种内源激素和4种多胺含量的动态变化。结果显示，雌花和两性花发育初期和后期GA₃含量高于雄花；多数时期雄花ZT和IAA含量高于雌花和两性花；雄花发育中、后期 ABA含量较大幅度上升，且明显高于雌花，两性花ABA含量变化趋势与雌花相近，但整体明显高于雌花；ZT/GA₃比值雄花高于雌花和两性花。由此表明，较高的GA₃含量，较低的ZT和IAA含量有利于雌蕊的发育，较高的ABA含量有利于雄蕊中、后期的发育。花蕾发育初期，尸胺(Cad)含量从高到低依次为雌花、两性花和雄花，随着发育的进行，雄花和两性花中Cad含量呈现较大幅度上升，而雌花始终保持在相对较低的水平。因此，相对较高的Cad含量可能有利于早期雌蕊的发育，较低的Cad含量有利于雄蕊的发育，而后期则相反，高含量的Cad有利于雄蕊的成熟；高含量的Spd可能有利于早期雌蕊的发育。     

甜瓜花器官发育相关基因的电子克隆及表达分析

以厚皮甜瓜（Cucumis melo）‘WI998’（纯雌系）与‘TopMark’（雄全同株）配制杂交组合获得的RIL₆群体为材料,对分离后代雌雄异花同株,雄全同株,完全花植株及纯雌株进行转录组测序,利用生物信息学分析方法,获得花器官发育相关基因序列Un16008,命名为CmTs2。该序列位于甜瓜第12条染色体上,长度1 842 bp,119 ~ 832之间存在一个包含237个氨基酸的最大开放式阅读框,成熟蛋白分子量为25 358.8 D,该蛋白不稳定系数为21.6,脂肪系数是99.11,平均亲水系数为0.078。通过Blast比对发现该基因与多种作物Tasselseeds2（TS2）基因具有高度同源性,其中与黄瓜Tasselseeds2基因同源性高达96%。通过实时荧光定量PCR检测,发现该基因在甜瓜雌雄异花同株花和叶片中表达量均高于其它性别类型植株,在纯雌株中表达量最低,说明该基因可能参与甜瓜花器官发育,尤其是雌花发育的过程。     

Genetics and molecular mapping of gynoecious (F) locus in cucumber (Cucumis sativus L.)

Gynoecious is an important economic trait of cucumber for determinant of earliness and yield, yet genetic mechanism is not well understood for this trait. The experiment was conducted using F₂ mapping population by crossing of PPC-2, a gynoecious and parthenocarpic line with Pusa Uday (monoecious and non-parthenocarpic cultivar). Out of 179 SSR markers screened, 39 markers differentiated the gynoecious and monoecious parents. However, only 17 markers were segregating with F₂ mapping population, those were used for genotyping and linkage map analysis and these markers were placed along with F locus on chromosome 6 covering a total distance of 100.4cM. The SSR markers, SSR13251 and UW020605 were found to be closely linked to gynoecious (F) locus at 1.0 and 4.5 cM, respectively. The segregation of F₂ population of PPC-2 × Pusa Uday and GPC-1 × Punjab Naveen and test crosses for sex type herein suggested that single dominant gene controlled the gynoecious sex expression in cucumber particularly in gynoecious genotypes PPC-2 and GPC-1. Therefore, the monogenic dominant nature of gynoecious sex identified in the present experiment and SSR markers closely linked to the F locus will be useful in marker-assisted backcross breeding for transfering gynoecious trait into horticulturally desirable varieties.     

Growth and abortion of flower buds in gynoecious cucumber plants in relation to temperature

The growth of flower buds in a gynoecious cucumber cultivar was studied at 15, 21 and 27°C. Competition between buds in the same node was measured as the ratio of the growth rates of individual buds. This competition was stronger at 21 and 27 than at 15°C. At 15°C, at least 3 buds were about equally strong; at 21°C, 2 buds developed fast and suppressed the other buds; at 27°C, the first bud which developed suppressed all other buds. When plants were transferred from one temperature to another, the growth-rate ratio between buds in older nodes was maintained. In nodes formed in the new temperature regime, the growth-rate ratio was determined by the new temperature.     

Sex inheritance and development of gynoecious hybrids in bitter gourd (Momordica charantia L.)

Gynoecism has been used on a large scale in heterosis breeding of cucumber. Production of hybrid seeds in bitter gourd is highly expensive because it is done mainly through hand pollination. But utilization of a gynoecious line would be more economical and easier method. Since gynoecious parent has only female flowers, the open pollinated seeds it produces will be F₁ hybrid. It will reduce the cost of male flower pinching and hand pollination. Two gynoecious lines, DBGY-201 and DBGY-202 were crossed with two monoecious cultivars, “Pusa Do Mausami” and “Pusa Vishesh” and their F₂ populations were observed to determine the inheritance pattern of gynoecious sex form. Another experiment was conducted to determine magnitude of heterosis by utilizing nine inbred lines including one gynoecious line, DBGY-201 in diallel (without reciprocal) mating system. All the gynoecious hybrids showed significant heterosis in a desirable direction for traits like sex ratio (male:female), days to first picking, number of fruits per plant, yield per plant, and vine length. The segregation pattern in F₂ suggested that gynoecious sex form in bitter gourd was governed by single recessive gene. The gynoecious hybrids DBGY-201 × “Pusa Vishesh” and DBGY-201 × “Pusa Do Mausami” were important for early harvest (～50 days after sowing) whereas DBGY-201 × “Priya” and DBGY-201 × “Arka Harit” were promising for higher yield.     

黄瓜全雌性基因连锁的AFLP和SCAR分子标记

 本研究以全雌品种‘戴多星’自交系和弱雌品种‘北京截头’自交系为双亲杂交获得F₁ ,然后得到F₂ 性型分离群体, 利用分离群体分组分析法(Bulked Segregant Analysis, BSA) 构建全雌和弱雌两个基因池, 筛选了64对AFLP选择性引物EcoR I-NN +Mse I-NNN组合, 发现EcoR I-TG +Mse I-CAC引物组合在全雌基因池中扩增出一条分子量为234 bp的特异带。经F₂ 代单株验证, 该特异条带能在全雌单株中稳定出现。以MAP MAKER (Version 310) 软件分析, 该标记与全雌性位点的连锁距离在617 cM。命名该连锁标记为TG/CAC₂₃₄。将该特异条带回收、克隆、测序, 设计特异SCAR引物, 再对F₂ 代单株基因组DNA进行扩增, 仅在全雌单株中扩增出1条分子量为166 bp 的特异带, 表明已成功地将与黄瓜全雌性连锁的AFLP标记转化为操作简便、表现稳定的SCAR标记, 该标记命名为SA₁₆₆。     

Monoecy and gynomonoecy in Spinacia oleracea L.: Morphological and genetic analyses

Two cosexual spinach lines, 03-336 and 03-258, were examined to investigate their sex expression. We found that line 03-336 consisted of plants bearing both pistillate and staminate flowers on the same individual (strictly monoecious type), while line 03-258 consisted of plants with both pistillate and hermaphroditic flowers on the same individual (gynomonoecious type). Interestingly, the two lines exhibited distinct patterns of inheritance when crossed with the pistillate plants derived from a dioecious line. The strictly monoecious character was considered to be conditioned by the known allele X^m, whereas the gynomonoecious character appeared to be governed by a novel monoecious allele or modifying gene(s) that may affect the sex expression.     

The effect of chilling on the termination of rest and flower bud development of the chinese gooseberry

Chinese gooseberry (Actinidia chinensis Planch.) cuttings were collected from two pistillate cultivars and one staminate clone at regular intervals throughout the winter, and chilled for various periods up to 50 days, before being forced into growth.The Chinese gooseberry had a low chilling requirement for bud break although chilling enhanced this process. Flower evocation occurred at least two months prior to leaf fall and before natural chilling commenced. Chilling, however, enhanced floral development by preventing the premature abortion of flower buds. There was a differential chilling response between the pistillate cultivars and the staminate clone.     

The modification of sex expression in papaya (Carica papaya L.)

Field trials with six times normal nitrogen supply produced a significant increase in the proportion of female papaya plants. The addition of nitrogen to the soil increased the leaf nitrogen content but had no effect on the amino acid content. Leaves of male plants contained more nitrogen than female plants but no difference could be detected in the contents of amino acids in the leaves and flower buds. Foliar application of 1-naphthalene acetic acid (NAA) and 2-chloroethyl-trimethyl ammonium chloride (CCC) resulted in a higher female : male ratio, but maleic hydrazide (MH) and gibberellic acid (GA₃) enhanced maleness. The animal sex hormone stilboesterol dipropionate had no apparent effect on sex expression but the application of testosterone propionate increased the proportion of male plants.     

黄瓜MADS-box基因家族生物信息学分析及CsMADS-box AGL62的克隆

分别以全雌系D1104-2-4、强雌系D1158♀-2、雌雄异花同株D0528-2为母本,与1个雌雄异花同株父本D0708-2杂交获得杂种一代,调查开花初期、根瓜期、盛果期3个时期雌性相关农艺性状,筛选出差异显著的组合进行转录组测序分析进而分析黄瓜杂交组合间MADS-box家族的差异基因。结果表明:以全雌系D1104-2-4为母本的杂种一代C15-114在雌花数、雌花节率上均具有超亲优势,3个杂交组合F1均具有早熟的中亲优势。转录组分析结果发现,以全雌系D1104-2-4为母本的杂种一代基因表达情况显著的偏向母本,挑选的37个黄瓜MADS-box家族基因只有6个与母本有表达差异,17个与父本有表达差异。生物信息学分析并命名了1个差异最显著的基因Cs MADS-box AGL62,该基因全长745 bp,编码187个氨基酸,编码的蛋白属于线粒体中不稳定的亲水性跨膜蛋白,NCBI比对结果表明该蛋白是1个转录因子,系统进化分析发现其与甜瓜中AGL62同源性最高,在黄瓜中该蛋白和其他黄瓜AGL62亚型属于同一分支并且属于I型。     

甜瓜雌雄异花同株杂交后代雌花率的遗传分析

利用雌雄异花同株品系3-2-2和雄全同株品系TopMark为亲本,配置杂交组合,获得6世代群体(P1,P2,F1,F2,BC1P1,BC1P),调查定植后第1朵雌花开放到定植后60 d内的雌花率,通过主-多基因混合模型分析其遗传效应.结果表明:甜瓜雌雄异花同株植株杂交后代雌花率受到1对主效基因控制,同时存在2对微效基因,主基因遗传率为83.33％,微效基因的遗传率为8.125％,同时讨论了利用雌花率分析雌雄异花同株遗传规律的可行性.     

西瓜3种性型花器官性别分化的细胞形态学观察

以西瓜3种性型(雌雄异花同株系、全雌系和完全花雄花系)的雄花、雌花和完全花3种花芽为材料,制成石蜡切片,进行西瓜花芽不同发育时期的细胞形态学观察。结果表明:西瓜3种性型之间性别分化起始及同种性别之间花器官发育过程并没有明显的形态学差异。雄花、雌花和完全花3种花芽分化首先要经过一个两性期,即雄蕊原基和雌蕊原基同时存在时期,在两性期之前三者不存在明显的形态学差异;当花芽长度为0.8~1.0mm时两性期结束,开始雄花、雌花和完全花的差异发育,此后雄花和完全花的雄蕊原基快速增大,雌花和完全花的雌蕊原基开始分化为柱头和子房。在3种花芽发育过程中,完全花的花芽在两性期后明显比同时期的雄花、雌花的花芽长度更长。全雌系中的"畸形雌花"出现在两性期后,雌蕊原基的凹陷发育过程受阻,而本该停止发育的雄蕊原基继续分化,最终发育成可产生少量成熟花粉粒的雄蕊。     

‘藤稔’葡萄冬季休眠后期花芽发育相关基因表达的分析

 以6 年生‘藤稔’葡萄（Vitis vinifera L.‘Fujiminori’）为试材,在枝条不同节位进行短截 处理,对9 个花发育相关基因的时空表达进行研究。结果表明,在冬季休眠后期虽然VvAP1、VvAP2、VvAP3、 VvAG、VvFUL、VvSOC1、VvLFY 和VvFLC 基因的相对表达水平较低,但冬芽仍可进行花芽分化;短截 处理可以促进花发育,增加各节位芽中基因的相对表达量;同一枝蔓上的中部芽比上部芽和下部芽发育 好,基因相对表达量高,花芽生长发育时间长。     

GA_3、CEPA对板栗内源碳水化合物及蛋白质含量的影响

在自然状态下，随着雌花的分化进程，板栗上部芽表现出蛋白质和蔗糖的积累，同时伴以淀粉和还原糖的消耗，下部芽的变化则不明显，其中蔗糖反而呈下降趋势；GA3处理增加了蛋白质和蔗糖的积累而加速了还原糖和淀粉的消耗，CEPA处理使还原糖和淀粉消耗减缓的同时也阻碍了蛋白质和蔗糖的积累。     

西瓜核型雄性不育两用系POD同工酶研究

用PAGE(聚丙烯酰胺凝胶)电泳技术,研究了西瓜核型雄性不育两用系不同器官的POD(过氧化物酶)同工酶酶谱差异。试验结果表明,不育株与可育株主要差异在雄花蕾,表现为不育株较可育株多2条活性强的酶带;其他器官差异小,仅表现为酶谱带强弱不同。并对POD同工酶与雄性不育性的相关性进行了研究。     

钝叶柃不同性别植株花期叶片内源激素含量的变化

柃木属(Eurya Thunb.)植物为严格的雌雄异株植物,性别变异现象极为少见。但作者野外调查发现,钝叶柃(Eurya obtusifolia H.T.Chang)存在显著的性别变异现象。以钝叶柃雌株、雄株和两性株花发育过程中关键时期的叶片为材料,采用高效液相色谱法对生长素(IAA)、赤霉素(GA_3)、玉米素核苷(ZR)和脱落酸(ABA)的含量进行测定和比值比较,并分析其动态变化规律。结果表明:在钝叶柃3种性别植株花发育过程中,叶片中GA_3和ZR含量增加有利于雄花和两性花的发育,高水平IAA有明显的促雌效应,ABA对性别分化的影响还不确定。在激素平衡中,ABA/ZR、IAA/ZR、GA_3/ZR比值的增加对雌花的发育有促进作用,而较高的ABA/IAA、GA3/IAA比值更有助于雄花的发育。