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.     

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。     

Vegetative and floral bud abortion in cucumber plants: Hormonal and environmental effects

The acropetal, zonal pattern of axillary buds along the main shoot of the cucumber plant, which includes bare nodes (no appreciable development), staminate, mixed and, lastly, pistillate (or hermaphrodite) ones, is described in monoecious, gynoecious, andromonoecious and hermaphrodite genotypic lines grown at various seasons and treated with gibberellin (GA_{4 + 7}) or with an ethylene-releasing compound (CEPA, 2-chloroethyl phosphonic acid).Under optimal growing-conditions, GA given to gynoecious and hermaphrodite plants mimicked summer conditions by increasing the number of basal bare nodes and the proportion of abortive female buds and staminate buds. CEPA given to monoecious and andromonoecious plants generally imitated winter conditions by decreasing the proportion of bare and abortive nodes, and increasing female tendency.Bud abortion was also noticed under sub-optimal growing-conditions (low light intensities and relatively low temperatures) during the winter. This seems to be physiologically unrelated to the above mentioned phenomena.     

苦瓜遗传育种研究进展

本文从苦瓜果实性状遗传、雌性系育种及性别分化、抗病育种、单倍体育种以及新品种选育方面,综述了苦瓜遗传育种的研究进展,并针对我国苦瓜育种面临的问题对未来发展趋势进行了展望。     

Development of novel EST-SSR markers for cucumber (Cucumis sativus) and their transferability to related species

We report on the development of novel simple sequence repeat (SSR) markers from publicly available Cucumis sativus expressed sequence tags (ESTs) and on their transferability among related species. In total, 533 di- to penta-type SSRs were identified from 6344 cucumber ESTs retrieved from GenBank. Identified SSRs mainly comprised of di- and tri-nucleotide repeats, of which AG and AAG motifs were much abundant. A total of 392 SSR-containing unigenes (non-redundant ESTs/consensus sequences) were suitable for primer design. From these, 35 primer pairs were designed as representative samples and 28 were usable markers. Twenty-six out of 28 usable markers revealed polymorphism among 21 cucumber accessions with 2–7 alleles detected (mean = 3.77) and their polymorphism information content (PIC) values ranged from 0.091 to 0.748 (mean = 0.388). The polymorphism observed herein partially arose from the null alleles which occurred at the multiple homoeoloci detected by the markers. Transferability of the 28 EST-SSR markers was investigated in four other cucurbits: melon, watermelon, pumpkin and gourd which showed frequency of 92.9%, 57.1%, 53.6% and 60.7%, respectively. The EST-SSR markers developed herein will complement the currently available genomic SSR markers and may be useful for genetic studies in cucumber and related species.     

枳根cDNA全长文库的构建与分析

为了解柑橘优良砧木枳（Poncirus trifoliata）根部的基因表达信息,利用SMART技术构建了枳的根组织全长cDNA文库。检测结果显示所建枳根原始文库的容量为1.08 × 106 cfu ? mL-1,扩增后文库容量为1.30 × 109 cfu ? mL-1,重组率为95%。通过文库随机测序获得182个长度大于100 bp的ESTs序列（GenBank登录号为JK316116 ~ JK316297）,序列拼接后得到96个Unigenes,包括12个Contigs和84个singletons。其中,68个Unigenes可与GenBank中登录的基因相匹配,56个与已知的柑橘基因相匹配,36个为全长基因。对24个全长基因完成了功能注释,并进一步开展了生物信息学分析,发现有6个应激反应基因和3个根发育相关基因。     

苗期喷施乙烯利对4种雌雄同株异花瓜类制种产量的影响

为了测试乙烯利对不同雌雄同株异花瓜类制种产量的影响,以瓠瓜、苦瓜、南瓜和黄瓜作为材料,在2叶1心和3叶1心这2个时期分别喷施100、150、200、250、300 mg/L乙烯利。结果表明:喷施适宜浓度的乙烯利可显著降低4种瓜类株高,有效降低第1雌花节位和增加20节内雌花数量,增加第1朵雌花开花所需时间,但对单瓜种粒数影响较小。当乙烯利使用浓度为瓠瓜150 mg/L、苦瓜200 mg/L、南瓜200 mg/L和黄瓜100 mg/L时,4种瓜类制种综合增产效果最佳。     

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。     

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。     

紫背天葵高通量转录组测序分析

为探讨紫背天葵(Gynura bicolor)花青素等物质合成的遗传基础,采用高通量测序技术(Illumina HiSeq 2500)对其嫩叶进行转录组测序,共获得21 387 624个序列读取片段(reads),将测序数据进行序列组装后,获得33 314个单基因簇(Unigene),其中超过1 kb的7 792个Unigene中共检测到2 387个SSR位点。对所得Unigene进行不同数据库注释,22 048和14 417个Unigene分别在Nr和Swiss Prot数据库有同源比对信息,并发现有29个Unigene与花青素合成相关;Pfam功能注释到13 909个Unigene分为5 198类相关蛋白功能区域,有12个Unigene涉及到花青素合成;GO注释到11 613个Unigene分为细胞组分、分子功能及生物学过程等3大类51个功能组,有24个Unigene与花青素合成有关;COG注释到的6 589个Unigene功能系统分为24类,而KOG注释到的13 498个Unigene功能系统分为25类;以KEGG数据库为参考,将4 466个Unigene定位到108个代谢途径分支,其中有47个Unigene与类黄酮生物合成相关。     

全雌系苦瓜两性花分化的化学调控

以不同苗龄全雌系苦瓜X黑-d-d为试材,研究硝酸银不同浓度、不同喷施次数对其诱雄的效果。结果表明,硝酸银诱导全雌系苦瓜X黑-d-d两性花分化的关键时期是三叶期,300mg·L-1硝酸银喷施2次,每次间隔5d的诱雄效果最好。     

Molecular characterization of an S-adenosylmethionine decarboxylase gene from floral organ differentiating axillary bud in calamondin (× Citrofortunella mitis)

Summary

Calamondin (× Citrofortunella mitis J. Ingram & H. E. Moore) is known for its ability to develop floral organs to flower in the adult phase during the four seasons. To provide information on molecular events during floral differentiation from axillary buds, we constructed a cDNA library from floral differentiating axillary buds of adult phase calamondin. Ninety-six cDNA clones were randomly selected from this library and sequenced. Ninety-six nonredundant ESTs were identified. Two clones encoded S-adenosylmethione decarboxylase, a key enzymes in polyamine biosynthesis. CmSAMDC 1 (Citrofortunella mitis s-adenosylmethionine decarboxylase 1) contained an 1083 bp ORF that encoded a putative SAMDC precursor of 361 amino acids. Northern blot analysis revealed that CmSAMDC 1 was expressed in axillary buds prior to floral differentiation and in axillary buds immediately after floral differentiation, in immature flower (5 mm), in fully developed flowers (120 mm long), in floral parts (petals, pistils, and stamens) of trees in the adult phase of development, but not in leaves or axillary buds of juvenile phase nucellar seedlings or leaf tissue from trees in adult phase of development. In situ hybridization revealed expression of the CmSAMDC 1 gene in the floral apex of axillary buds after differentiation, and in the phloem of axillary buds and petioles. These results suggest that SAMDC could play a role in actively differentiating and developing reproductive and vegetative organs.     

苦瓜新品种‘丰绿3号’

‘丰绿3号’苦瓜是以强雌系B201-b1作母本,优质自交系E12206-e6作父本配制而成的杂交一代新品种。果实长圆锥形,深绿色,条瘤明显,长25 cm左右,横径约6.2 cm,肉厚1.0 cm,单瓜质量约440 g,商品性好。从播种至始收春季76 d,秋季54 d。单株坐果力强,丰产性好,平均产量为36.61t·hm~(-2)。耐热、耐涝性强,植株不易早衰。适宜华南地区春、秋季和海南冬季种植。     

黄瓜抗白粉病品系cDNA文库构建及EST分析

 采用SMART技术构建了高抗白粉病的黄瓜品系JIN5-508的叶片cDNA文库,并对其进行了质量鉴定。结果表明,该文库的重组率为95.3%,库容量为1.02 × 106 pfu · mL-1,平均插入片段为0.5 kb左右,是一个高质量的cDNA文库。利用该文库随机挑选的8 352个克隆从5′ 端进行单向测序,共获得8 035个有效的ESTs（expressed sequence tags）,序列的平均长度为838 bp。进一步的生物信息学分析表明：EST序列拼接出3 467个unigenes,其中包括953个contigs和2 514条singletons,文库冗余度较低,为56.9%;从文库中筛选出了大量的低丰度表达基因,约占unigene总数的72.5%,说明文库的复杂性较高;1 991个unigenes获得分子功能注释,其中与蛋白结合活性和催化活性相关的基因分别达到了41.34%和38.72%;在获得功能注释的unigene中,有部分抗病抗逆相关基因高丰度表达,其中3个unigenes与拟南芥白粉病抗性基因Mlo2、Mlo8和Mlo14高度同源,表明该文库可为进一步研究黄瓜抗白粉病相关的特异基因的克隆及功能验证奠定基础。     

苦瓜第1雌花节位和雌花数的杂种优势

将8个绿苦瓜高世代自交系分为两组;共配制15个杂交组合,对第1雌花节位和主蔓前期雌花数的杂种优势进行了研究。结果表明:第1雌花节位有6个组合表现负向离中优势,有13个组合表现为负向越亲优势,且有4个组合低于小值亲本;雌花数有5个组合表现为正向离中优势,有4个组合表现为正向超亲优势。通过对第1雌花节位与雌花数两个性状亲子关系的讨论提出实现早熟性和晚熟性育种目标的亲本选配方法。     

鸟巢蕨转录组高通量测序及分析

采用新一代高通量测序技术Illumina HiSeq 2000对鸟巢蕨转录组（Asplenium nidus）进行测序,共获得29 254 595个序列读取片段（reads）,包含了5 908 586 517个碱基序列（bp）信息。对reads进行序列组装,共获得42 907个单基因簇（Unigene）,平均长度936 bp,序列信息达到了40.16 Mb。数据库中的序列同源性比较表明,24 993个Unigene与其他物种的已知基因具有不同程度的同源性。鸟巢蕨转录组中的Unigene根据GO功能大致可分为细胞组分、分子功能和生物学过程3大类51个分支,其中有大量的Unigene与代谢进程、结合活性、催化活性和细胞进程相关。将Unigene与COG数据库进行比对,根据其功能大致可分为24类。KEGG数据库作为参考,依据代谢途径可将Unigene定位到116个代谢途径分支。SSR位点查找发现,从42 907个Unigene中共找到6 067个SSR位点。SSR不同重复基序类型中,出现频率最高的为AG/CT,其次是AC/GT、A/T和AGG/CCT。针对这些序列,设计了20对引物进行了扩增效率和多态性检测,其中7对引物在不同蕨类材料中表现出多态性。     

基于银杏花芽3个分化时期转录组测序的相关基因筛选与表达分析

鉴定银杏花芽分化调控的关键基因,揭示银杏花芽分化调控的主要分子机制,为缩短银杏童期和选育银杏早花品种提供理论指导。本研究中采用高通量测序技术对银杏花芽分化3个时期（花芽未分化期、花芽分化始期、花芽分化盛期）的样品进行转录组测序,并分析数字表达谱,筛选开花调控相关基因并进行荧光定量PCR（RT-qPCR）表达验证。转录组测序共产生27.52 Gb原始数据,注释到8大功能数据库（GO、COG、KEGG、KOG、NR、Pfam、Swiss-Prot、eggNOG）上的 unigene 总数为35 179个。通过GO分类和KEGG Pathway 富集性分析,将unigene分别归于55个GO类别和126个代谢途径。差异表达基因分析显示,花芽未分化期较花芽分化始期有2 253个基因上调,2 032个基因下调;花芽分化始期较花芽分化盛期有1 770个基因上调,1 901个基因下调;花芽未分化期较花芽分化盛期有1 865 个基因上调,2 042个基因下调。发掘出大量的开花相关的基因涉及5个开花调控途径（光周期途径、春化途径、赤霉素途径、自主途径和年龄途径）。筛选出gene.Gb_17618（GI序列）、gene.Gb_19790（FT/TFL1序列）、gene.Gb_16301（AG序列）、gene.Gb_28337（花发育MADS-box序列）、gene.Gb_01884（SOC1序列）和gene.Gb_41704（CO序列）等6个银杏花芽分化差异表达关键基因序列,荧光定量PCR检测表达水平与转录组结果一致。     

苦瓜种子大小和单粒质量的遗传分析

以苦瓜自交系K7-359(P_1)和K7-422(P_2)分别作为母本和父本,通过杂交获得F_1,F_1自交获得F_2群体,采用主基因+多基因混合遗传模型分析苦瓜种子长度、宽度和单粒质量的遗传规律。结果表明,苦瓜种子长度和单粒质量的遗传均符合加性-显性-上位性多基因遗传模型(C-0模型),多基因遗传率分别为84.91%和76.55%,说明对苦瓜种子长度和单粒质量的选择宜在高世代进行;苦瓜种子宽度的遗传符合1对加性-显性主基因+加性-显性-上位性多基因模型(D-0模型),主基因和多基因遗传率分别为79.30%和3.86%,主基因遗传效应主要以加性效应为主,说明对种子宽度的改良可以采用组合育种的策略,且适宜在早期世代进行选择。