QTL analysis of early flowering of female flowers in zucchini(Cucurbita pepo L.)

Early flowering promotes early maturity, production, and the capacity to counteract biotic and abiotic stresses, making it an important agronomic trait in zucchini. The present study demonstrated that the zucchini inbred line ‘19’ consistently flowered early, taking significantly fewer days to bloom the first female flower(DFF) than the inbred line ‘113’. Genetic analysis revealed that DFF, an inheritable quantitative trait, is controlled by multiple genes. Based on the strategy of quantitative trait locus(QTL) sequencing(QTL-seq) combined with linkage analysis, three QTLs for DFF were identified on chromosomes 4, 11, and 20. This study used additional F2 populations grown under different environmental conditions for QTL mapping analysis of DFF with insertion/deletion(InDel) markers to validate these results. Using the composite interval mapping(CIM) method of R/qtl software, we only identified one major locus under all environmental conditions, located in a 117-kb candidate region on chromosome 20. Based on gene annotation, gene sequence alignment, and qRT-PCR analysis, we found that the Cp4.1LG20g08050 gene encoding a RING finger protein may be a candidate gene for the opposite regulation of early flowering in zucchini. In summary, these results lay a foundation for a better understanding of early flowering and improving early flowering-based breeding strategies in zucchini.

黄瓜和南瓜Bcl-2相关抗凋亡家族全基因组鉴定与表达模式分析

【目的】分析Bcl-2相关抗凋亡(Bcl-2 associated athanogene,BAG)家族蛋白成员在黄瓜和南瓜中响应非生物胁迫以及在黄瓜/南瓜嫁接愈合过程中响应光照的表达模式,为解析黄瓜/南瓜嫁接苗嫁接愈合机理及黄瓜和南瓜等蔬菜的抗性分子育种提供有利基因。【方法】基于黄瓜和南瓜基因组信息,利用生物信息学手段,对黄瓜和南瓜中BAG基因家族进行鉴定,并对其理化特性、染色体定位、基因结构、系统发育和共线性进行了分析。基于公共数据库及黄瓜/南瓜嫁接苗在嫁接愈合过程中转录组测序数据,分析BAG基因在黄瓜和南瓜中响应非生物胁迫以及在嫁接愈合过程中响应光照的表达模式。【结果】在黄瓜和南瓜中分别鉴定到12和18个BAG家族基因,均分为2个亚族,基因成员保守性高,I亚家族的BAG主要参与基因调控和逆境响应,而II亚家族的BAG主要参与植物的发育过程。黄瓜和南瓜BAG基因分别与拟南芥、水稻、番茄存在多种线性关系,但CsaV3_1G017210与拟南芥、水稻、番茄和南瓜中的BAG基因均不存在线性关系。不同BAG基因具有组织特异性表达模式。CsaV3_6G000970和CmoCh08G008520(BAG family molecular chaperone regulator 6)在黄瓜和南瓜响应非生物胁迫以及黄瓜/南瓜嫁接苗嫁接愈合过程中均发生上调表达。【结论】BAG家族基因在黄瓜和南瓜对非生物胁迫的响应以及黄瓜/南瓜嫁接苗嫁接愈合过程中对光的响应中具有差异性,协同调控了黄瓜和南瓜的生长发育及嫁接愈合,在黄瓜和南瓜非生物胁迫以及黄瓜/南瓜嫁接苗嫁接愈合过程中发挥着重要作用。

In vitro Regeneration of First-generation Inbred Progenies of Cucurbita pepo L. Double Haploids

Using seeds of inbred progenies of Cucurbita pepo L. DH12-11409 double haplolds as experimental materials, the effects of different hormone combinations, explant types and seedling ages on adventitious bud regeneration and rooting of C. pepo L. were investigated. According to the results, inoculating cotyledonary nodes of yellow-green cotyledons from 5-d-old C. pepo L. double haploids to MS ＋ 30 g/L Suc ＋ 8 g/L Agar ＋ 2.0 mg/L 6-BA ＋ 0.1 mg/L NAA led to the best results with the induction frequency of 90.0% and differentiation coefficient of 8.5. MS medium with addition of 0.05 or 0.1 mg/L NAA led to the highest rooting rate. Regenerated seedlings with 5 - 6 true leaves exhibited the highest survival rate of 90.0%, which was the optimal period for domestication and transplanting of regenerated seedlings. This study laid a solid foundation for high-efficiency utilization of heterosis of C. pepo L.

Use of Cucurbita pepo Oil to Fight against the UV Action on the Skin

Skin aging is a process most often attributed to UV [1] and also to the use of creams and other cosmetic products low in antioxidant compounds [2]. Photochemically stable pepo Cucurbita oil can be used as an exogenous cosmetic supplement due to its high antioxidant content. Incorporated in an agar, media containing a synthetic melanin solution with added pumpkin oil are subjected to UV light, the aging thus modeled is followed by the measurement of photoresistance values correlated with chemical and spectrophotometric analyses. This study confirms that pumpkin oil is highly effective in protecting the skin, especially the most sensitive skins such as babies’ skin [3] by reinforcing the action of melanin and also that of albinos without melanin. Indeed its SPF (Significant Sun Protection Factor) index estimated during this work is very consistent, i.e. more than 22% of UVB (280 - 315 nm) radiations are suppressed.

Study the Effect of Different Levels of Zucchini (<i>Cucurbita pepo</i>L.) on the Biological Indicators for the Prevention of Cardiovascular Disease in Rats Fed High-Fat Diets

The effects of fresh zucchini on nutritional status, and biological indicators for the prevention of cardiovascular disease in rats fed high-fat diets investigated in this study. Thirty Sprague Dawley rats were randomly divided into two main groups the first, negative control group Co (-) (n = 6), fed basal diet, The second group (n = 24) fed high-fat diet (containing basal diet + 5% tallow + 1% cholesterol + 0.02% bile salt). This group was divided into four subgroups each group 6 rats: group positive control co (+) fed high-fat diet only, group 1 (G (1)) fed high-fat diet plus 10% zucchini, group 2 (G (2) fed high-fat diet plus 15% zucchini and group 3 (G (3)) fed high-fat diet plus 20% zucchini. The levels of serum total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) were measured after eight wk. of experimental treatment. The pathologic changes of the heart, spleen and kidney were evaluated. SPSS, one way ANOVA was used to analyze the results. The results indicated that the mean values of body weight gain (BWG) and feed efficiency ratio (FIR) in G (2) and G (3) showed significant decrease compared to co (-), co (+) and G 1. The results showed that in groups (G1, G2 and G3) the relative weight of heart had significant increase when compared with control negative group. But liver and spleen weight had no significant difference when compared with control negative and positive group, the levels of TC, TG and LDL-C were significantly increased in co (+) (P < 0.05) Compared to co (-), all groups fed on high fat diet containing different levels of zucchini (10%, 15% and 20%) had significant decrease in TC and LDL-C compared with co (+), also G2 and G3 had improve significant in HDL-C when compared with co. (+). The groups fed on zucchini give results similar to group co. (-). The best treatment was zucchini (15% and 20%) which had lowest values of total lipid cholesterol and LDL-C, and the best values of HDL-C, HDL-C/TC % for all groups fed on high fat diet and zucchini increased non significantly (p ≤ 0.05) the HDL-C/TC % index compared to group co. (+). while, G2 and G3 gave significant decrease in LDL-C/HDL-C ratio compared to group co. (+). Morphologic changes of heart, spleen and kidney revealed that groups G2 and G3 had a similar preventive effect against CHD in this experimental model. In conclusion, results showed that zucchini had similar potential to attenuate CHD-related parameters in a mild oxidative stress induced by high-fat diet in rats.

The Effects of Simultaneous Application of Different Organic and Biological Fertilizers on Quantitative and Qualitative Characteristics of Cucurbita pepo L,

Understanding the relations and interactions between ecosystem components and plants is crucial for sustainable production of medicinal plants. To study the effect of simultaneous application of organic and biological fertilizers on yield and yield components of zucchini squash, split plot arrangement of factors based on randomized complete block design with three replications were used during 2009-2010 growing season. The mainplot factors were the type of organic fertilizers, including： （1） cow manure; （2） sheep manure; （3） chicken manure; （4） vermicompost; and （5） control. The subplot factors were the biofertilizes （Nitragin, containing Azotobacter sp., Azospirillum sp. and Pseudomonas sp.） utilization. The results showed the positive but non-significant effect of organic and biological fertilizers on yield and yield components of zucchini squash. Amongst the organic fertilizers, cow and chicken manure, have superiority compared to others. The highest seed oil and protein percent was obtained with application of chicken manure, however there was no significant difference between treatments in seed oil percent. The positive effect of organic and biological fertilizers on seed yield was higher than fruit yield. At a glance, application of cow manure solely was better than its application with nitragin. Nitragin application has no significant effect on some traits when utilized with sheep manure and vermicompost.

Direct Organogenesis from Cotyledonary Node Explants of Cucurbita pepo (L.)—An Important Zucchini Type Vegetable Crop

Cotyledonary node explants from 3 - 5-d-old seedling derived explants of Cucurbita pepo (L.) produced multiple shoots in Murashige and Skoog (MS) salts B5 vitamins containing medium in the presence of N6-benzylamino-purine BAP 1 mg/L + Thidiazuron TDZ 0.05 mg/L. After 4 weeks explants were subcultured to medium containing MS salts B5 Vitamins BAP 0.5 mg/L, Gibberellic acid GA3 1 mg/L and L-Glutamine 15 mg/L. Periodic excision of regenerated shoots from the explants increased shoot efficiency during subculture. The combination of L-Glutamine with BA and GA3 significantly increased the shoot proliferation. Elongated shoots were excised from shoot clumps and transferred to rooting medium containing indole-3-butyric acid (IBA, 1.0 mg/L). The rooted plants were hardened in small pot containing standardized garden soil, well developed plant transferred to greenhouse and survival rate was 85%.

Valorization of Cucurbita pepo Seed Oil in Soap Production

The present study was conducted with the aim of extracting and characterising Cucurbita pepo seed oil and then producing soap with important dermatological properties. The physico-chemical results of the Cucurbita pepo seed oil after extraction were satisfactory and indicate that the oil obtained contains long-carbonaceous fatty acids. The production of the soap was based on the realisation of a saponification reaction by reflux heating between the extracted oil and a solution of soduim hydroxyde 7M for one hour. The soap obtained is characterised by the following physico-chemical parameters: salt content, alkali content, pH, and humidity;which meet ISO 684-1974 standards. The antibacterial parameters of the soaps S1, S2, S3, S4, S5 and S6 were determined by the Agar Diffusion test using the well method and these soaps showed an interesting antibacterial activity against the germs pseudomonas aeruginosa and staphylococcus aureus. Soaps based on Raffia sese oil (kolo oil) and Elaesis guineensis oil (palm oil) have also been synthesized for comparison with soap obtained from Cucurbita pepo seed oil.

美洲南瓜(Cucurbita pepo)种皮苯丙氨酸解氨酶基因克隆与表达分析

【目的】克隆有壳美洲南瓜种皮PAL基因(CP-PAL),研究PAL基因在有壳和裸仁美洲南瓜种皮发育过程中的表达特性,为揭示美洲南瓜种皮发育机理及木质素积累在南瓜种皮发育中的作用等方面提供理论依据。【方法】利用RT-PCR,结合RACE技术克隆CP-PAL的全长序列并进行生物信息学分析;利用实时荧光定量PCR技术,采用2-△△Ct方法对种皮发育过程中PAL基因的表达进行分析。【结果】CP-PAL序列全长为1 720 bp,含有一个1 359bp的ORF,114 bp 5′端非翻译区、236 bp 3′端非翻译区及11 bp polyA结构,可编码452个氨基酸,分子量为48.86 kD,等电点为6.55,原子总数为6 885个,分子式为C2158H3449N607O657S14。通过BLASTX比对表明CP-PAL核苷酸序列及其氨基酸序列与黄瓜PAL核苷酸及其氨基酸序列的相似性最高。CP-PAL包含PAL-HAL、PLN02457及phe_am_lyase 3个结构域及酶活性中心序列(GTITASGDLVPLSYIA),属于Lyase_I_Like超家族。CP-PAL不具有导肽及信号肽,为非跨膜蛋白,可能定位于细胞质及内质网上,属可溶性蛋白。CP-PAL蛋白含有4个酪蛋白激酶Ⅱ识别位点、6个蛋白激酶C识别位点、12个豆蔻酰化位点及2个糖基化位点。此外,分析可知CP-PAL有18个丝氨酸磷酸化位点、6个苏氨酸磷酸化位点及5个酪氨酸磷酸化位点。无规则卷曲是CP-PAL蛋白二级结构中最大量的结构元件,α-螺旋和延伸链分散于整个蛋白质中,且N-末端以无规则卷曲形式存在,C-末端以延伸链形式存在。CP-PAL氨基酸序列同挑选的其他14种植物的PAL氨基酸序列进行多重序列比较,发现功能区域的氨基酸序列较为保守,N-端的差异最大。系统进化树分析表明CP-PAL和黄瓜PAL蛋白的亲缘关系最近。CP-PAL蛋白三级结构以α-螺旋为主要结构元件,β-转角和无规则卷曲较少。实时荧光定量PCR分析表明PAL基因在有壳和裸仁美洲南瓜种皮发育中呈现反向对应的变化趋势:有壳美洲南瓜种皮PAL基因在自交授粉20 d后表达量增加,而裸仁美洲南瓜种皮PAL基因在20 d后表达量下降。整个种皮发育过程中,PAL基因在裸仁美洲南瓜中的表达量低于其在有壳美洲南瓜中的表达量。【结论】从有壳美洲南瓜种皮中克隆得到与木质素合成相关的PAL基因,该基因可能通过参与调控种皮木质素的合成从而影响美洲南瓜裸粒品种的种皮发育。

美洲南瓜(Cucurbita pepo)种皮发育形态观察及其相关酶活性测定

采用组织整体染色和石蜡切片相结合的方法对美洲南瓜有壳品种04LAg-26-2和无壳品种04LAg-26-28种皮发育形态结构比较观察。结果表明,有壳美洲南瓜种皮是由表皮层、下皮层、厚壁组织层、软组织层和绿色组织层构成,无壳裸仁南瓜的形成主要是授粉后18 d种皮发育过程中厚壁组织中的细胞变形、瓦解,缺少木质素的积累而导致了下皮层、厚壁组织层的缺失;对2个品种的种皮、叶片、叶柄和瓜囊不同组织中木质素合成相关酶活性测定结果表明,无壳品种04LAg-26-28在授粉18 d后,种皮中苯丙氨酸解氨酶(PAL)、肉桂酸-4-羟基化酶(C4H)、4-香豆酸-辅酶A连接酶(4CL)、肉桂醇脱氢酶(CAD)和过氧化物酶(POD)活性明显降低,而叶片、叶柄和瓜囊中这几种酶活性在授粉18 d后,呈逐渐增加的趋势。

北方早春棚室西葫芦嫁接育苗栽培技术

为了提高西葫芦的产量和品质,满足市场需求,根据多年的从事棚室栽培技术经验,总结了北方棚室西葫芦嫁接育苗栽培技术,以供广大种植户参考。

硅对盐胁迫下西葫芦幼苗水分代谢及光合特性的影响

【目的】探究外源硅对盐胁迫下西葫芦幼苗水分代谢及光合作用的保护机制,可为西葫芦抗盐栽培提供理论参考。【方法】以西葫芦品种‘寒绿7042’为试材,通过水培试验,共设置对照、硅处理(0.3 mmol/L Na_(2)SiO_(3)·9H_(2)O)、盐胁迫(150 mmol/L NaCl)、盐胁迫+硅等4个处理,处理10 d后测定幼苗的根系形态和活力,叶片含水量、水势、光合气体交换参数、叶绿素含量、叶绿素荧光参数及根系质膜水通道蛋白基因的表达等指标。【结果】盐胁迫显著抑制了西葫芦幼苗根系的生长,降低了根系活力、叶片含水量、叶片水势和蒸腾作用,还破坏了植株的光合系统,导致净光合速率、气孔导度、叶绿素含量和PSⅡ光化学效率显著降低。外源硅显著改善了盐胁迫下西葫芦幼苗的根系形态,提高了根系活力,并通过促进根系质膜水通道蛋白基因PIP1;2、PIP1;3、PIP1;5、PIP1;7、PIP2;1、PIP2;4、PIP2;6、PIP2;8、PIP2;9和PIP2;12的表达改善植株体内的水分状况,提高叶片含水量和水势,增强蒸腾作用;外源硅还通过提高叶片气孔导度、叶绿素含量、净光合速率、PSⅡ最大光化学效率、PSⅡ实际光化学效率、光化学淬灭系数,降低胞间CO_(2)浓度和非光化学淬灭系数来增强盐胁迫下幼苗的光合作用。【结论】盐胁迫下添加0.3 mmol/L的硅可以有效改善西葫芦幼苗的水分代谢和光合作用状况,从而提高西葫芦幼苗抗盐胁迫能力。

NaCl胁迫对黑籽南瓜和西葫芦种子萌发影响的对比研究

用不同浓度的N aC l胁迫处理黑籽南瓜和西葫芦种子,观察其发芽情况,计算发芽率、发芽势、发芽指数和活力指数,进行对比分析。结果表明,黑籽南瓜种子萌发对盐水生境的适应性比西葫芦强,黑籽南瓜种子萌发的N aC l胁迫浓度适宜值为50.66 mm o l/L,而西葫芦为43.98 mm o l/L。

NaCl胁迫及Ca^(2+)和GA_3对南瓜属3种蔬菜种子发芽的影响

研究了NaCl胁迫对南瓜 (CucurbitamoschataDuch. )、笋瓜 (C. maximaDuch. )和西葫芦 (C. pepoL. )种子萌发的影响及不同浓度外源Ca2+和GA3对NaCl胁迫下南瓜种子发芽的效应。结果表明,用 30mmol·L-1 NaCl处理,南瓜种子的发芽率高于对照(蒸馏水),而用 100和 170mmol·L-1 NaCl处理,西葫芦和笋瓜种子的发芽率下降率为负值,表明较低浓度NaCl胁迫可一定程度提高西葫芦、笋瓜和南瓜种子的发芽率;高浓度NaCl胁迫对种子发芽有明显的抑制作用。种子萌发期耐盐能力从大至小依次为西葫芦、南瓜、笋瓜。在 170mmol·L-1 NaCl胁迫下,施加 5~30mmol·L-1外源Ca2+或浸种处理,对南瓜种子发芽有促进作用,但高浓度外源Ca2+ (≥50mmol·L-1CaCl2 )则具有抑制作用。在 170mmol·L-1 NaCl胁迫下,用不同浓度GA3浸种处理,对南瓜种子发芽有抑制作用。

美洲南瓜裸仁基因的AFLP和SCAR分子标记分析

以美洲南瓜有壳品系"金辉二号-2"和无壳品系"0516-2"进行杂交,获得193个F2单株作为作图群体,利用AFLP分子标记技术和集群分析法(BSA)对与美洲南瓜裸仁基因连锁的分子标记进行研究。结果表明,找到了与裸仁基因n连锁的4个AFLP标记:E19M60-C、E22M48-B、E13M60-A和E25M51-D,连锁遗传距离分别为8.9、14.1、17.5和18.8 cM。同时将E19M60-C转化为操作简便、表现稳定的SCAR标记,用于美洲南瓜的分子标记辅助育种。

白粉病菌侵染对美洲南瓜不同抗性品种PAL基因表达量的影响

为明确白粉病菌侵染对不同美洲南瓜抗性品种PAL基因表达量的影响,采用室内接种和qRTPCR技术分别测定不同美洲南瓜抗性品种接种苍耳单囊白粉菌后发病率和病情指数,并分析接菌后其不同组织部位PAL基因表达量的变化情况。室内测定结果表明,抗病品种小三星F2发病率和病情指数显著低于感病品种金12F2,尤其接菌后第13天,小三星F2发病率和病情指数分别为17.3%和15.6,而金12F2发病率和病情指数分别为93.3%和83.0。PAL基因表达量测定结果表明,不同抗性美洲南瓜接菌后,PAL基因相对表达量显著升高,且随着接种天数增加,其不同组织部位PAL基因相对表达量呈现出先增加后降低的趋势。抗病品种小三星F2接菌后叶片中PAL基因相对表达量增加较为显著,叶柄和茎秆次之,并且其相对表达量分别在第7和第9天达到最大值;感病品种金12F2接菌后叶片中PAL基因相对表达量较抗病品种低,但与对照相比,其PAL基因相对表达量显著增加,并分别在第9和第11天达到最大值。因此,PAL基因相对表达量与美洲南瓜抗白粉病强弱具有密切的关系。本研究可为南瓜抗病品种的选育提供科学依据。

ALA对低温胁迫下西葫芦幼苗光合特性的影响

为深入了解5-氨基乙酰丙酸(ALA)对西葫芦生长影响的光合生理机理,以‘早青一代’西葫芦为材料,研究ALA对低温胁迫下西葫芦幼苗光合特性的影响。结果表明,低温胁迫下,西葫芦幼苗叶绿素含量显著降低,净光合速率显著降低,单株叶面积和物质积累极显著减少;60mg/L的ALA可以减缓低温胁迫对西葫芦幼苗的影响,使总叶绿素含量增加29.96%,净光合速率(Pn)增加52.96%,叶面积增加39.80%,单株物质积累增加43.75%。60mg/L的ALA缓解低温胁迫对西葫芦幼苗的影响主要是通过叶绿素的增加和非气孔因素实现的。

美洲南瓜(Cucurbit pepo)WMV-2抗性遗传及分子标记研究

以美洲南瓜(Cucurbita pepo)抗病自交系‘0504’和感病自交系‘0223’为亲本杂交获得的157个F2单株为材料,研究南瓜对西瓜花叶病毒2号(WMV-2)抗性的遗传规律和分子标记。对亲本、F1及F2苗期接种WMV-2进行抗病性鉴定,结果表明,美洲南瓜对WMV-2的抗性由单隐性基因控制。采用集团分离分析法(BSA)和SSR技术,筛选出与WMV-2抗性基因连锁的标记CMTm157和CMTm158,遗传距离分别为10.9cM和6.2cM,其可以作为南瓜抗WMV-2育种的辅助选择分子标记。

番石榴实蝇和瓜实蝇的线粒体L-rRNA基因测序及双翅目昆虫L-rRNA基因的结构特征与系统发育关系分析

RNA二级结构比一级结构包含更多的信息,在物种系统发育分析中更能反应真实情况。因而,本研究对番石榴实蝇Bactrocera correcta和瓜实蝇B. cucurbitae的L-rRNA全基因进行了测序,构建了双翅目L-rRNA基因二级结构的模式图,并分析了其结构特征,而后基于H45～H47茎环结构参数和结构序列及L-rRNA结构序列对双翅目13科的系统发育关系进行了初步分析。结果表明:双翅目昆虫具有保守的L-rRNA二级结构;不同结构区碱基组成和分布不均匀,Ⅳ区和Ⅴ区的完全保守碱基含量最高,GC含量最高,绝大部分科Ⅵ区的AT斜率均小于0;科内特有碱基和科间少数保守碱基大部分为G或C。瘿蚊科与双翅目其他科具有较远的系统发育关系,而丽蝇科、寄蝇科和食蚜蝇科有相近的系统发育关系;虻科与网翅虻科同在一个小分支;蠓科与蚊科同在一个大的系统发育分支。利用单方面参数不易得到理想的进化结果,准确系统发育分析需要结合多方面的参数。

含聚天冬氨酸微生物肥料中养分比例对西葫芦生长的作用初探

针对国内温室西葫芦生产过程中存在的盲目施用肥料而造成的瓜菜产量和品质的降低问题进行了研究。以河北培根生物科技有限公司生产的微生物肥料(添加聚天冬氨酸)为供试肥料,以N、P_(2)O_(5)、K_(2)O不同养分比例设置试验处理(T1、T2、T3)、普通复合肥料(N 15%、P_(2)O_(5) 15%、K_(2)O 15%)作为CK处理进行了相关试验,通过对西葫芦不同生物阶段的多个指标进行综合分析,结果发现,在试验范围内T1(N 16%、P_(2)O_(5) 9%、K_(2)O 20%)处理效果最佳;所试验的肥料促进西葫芦的营养生长、促进开花坐果、加快果实发育速度,还能提高瓜菜外观品质、可溶性糖含量和V_C含量,显著提高了产量和经济效益。