A Good Seedless Ponkan Cultivar—Qianyang Seedless Ponkan

Qianyang Seedless Ponkan selected from common ponkan is a sport deviation cultivar without any seeds in its fruit. Matures in mid November, it has bigger avg.124.5g fruit, and the yield is 30 t ha-1 after 5 years' growing. There are no obvious in-year or off-year phenomena. Its fruits can store well without any treatments at room temperature for three or four months.

An Effective Method for Extracting Total RNA from Young Embryo of Seedless Litchi

A total RNA extraction method for young embryo of seedless litchi was introduced. CTAB, Phenol （saturated with water）, chloroform, Guanidine isothioeyanate were used as main extraction reagents. Polyphenolie compounds were removed effectively by added PVP into the extraction buffer solution. RNA was purified intensively by phenol, chloroform extraction, and ethanol deposition after deposited by LiCl. Both the results of formaldehyde denatured agarose gel eleetrophoresis and ultraviolet spectrophotometer analysis showed high integrity and purity of RNA. So the quality of extracted RNA could meet the demand of most molecular biology experiments that require higher quality RNA.

Flame Seedless葡萄胚珠、胚乳及胚发育与败育的研究

对Flame Seedless葡萄胚珠、胚乳及胚的发育和败育过程进行了系统的细胞学观察,并以有核葡萄品种北醇为对照对Flame Seedless的胚珠重量及纵横径进行了测定.结果表明:(1)Flame Seedless内、外珠被及珠孔发育异常,20%～30%的子房未出现受精现象,受精的胚珠重量和纵横径随着胚珠的发育,先呈增加趋势,在某一时期达到最高值,随后下降,最终退化为痕迹.(2)大约有30%的胚乳核能够进行正常分裂,形成胚乳组织,胚乳在花后30 d开始退化,其余胚乳核分裂异常.合子在盛花后第19天进行第一次分裂,经过二细胞原胚→多细胞原胚→球形胚,此后胚开始退化;也有的合子始终不发生分裂,最终退化.(3)由于胚乳分裂异常和提早退化,胚缺乏营养致使其发育中止和败育.(4)Flame Seedless胚挽救的最佳接种时期在花后第37天.

不同砧木对‘克瑞森无核’葡萄酚类物质含量及其合成相关酶活性的影响

以不同砧木嫁接的‘克瑞森无核’葡萄为试材,自根苗为对照,研究其果皮色泽、酚类物质含量以及其合成相关酶活性,以期通过砧木嫁接提高‘克瑞森无核’酚类物质含量,改善着色不均,进而提高果实品质。研究结果表明,砧木嫁接的成熟‘克瑞森无核’葡萄呈红色,砧木101-14、110R嫁接及自嫁接均降低L^(*)、b^(*),增加a^(*),而砧木SO4降低L^(*)、b^(*),同时砧木101-14、110R嫁接显著促进葡萄果实可溶性固形物的积累,降低其可滴定酸含量。砧木嫁接在不同程度上能促进‘克瑞森无核’葡萄的酚类物质的积累,成熟时砧木101-14、110R、SO4嫁接和自嫁接均能提高总酚、类黄酮和花色苷含量,其中砧木101-14嫁接的‘克瑞森无核’的总酚、类黄酮和花色苷分别较对照显著提高50.88%、52.00%和74.13%,促进效果最为明显。与对照相比,砧木101-14嫁接最有利于提高‘克瑞森无核’葡萄的酚类物质各合成相关酶活性,而砧木SO4嫁接对提高DFR和UFGT活性不明显。相关性分析表明,砧木嫁接苗果皮中酚类物质与DGR和UFGT活性相关性较高,其共同作用影响酚类物质的变化。研究发现,砧木嫁接促进‘克瑞森无核’葡萄果皮着色,砧木101-14、110R、SO4嫁接和自嫁接均能通过促进DFR和UFGT的活性,提高果皮中酚类物质含量,并以砧木101-14表现最好。

ClBBM and ClPLT2 function redundantly during both male and female gametophytes development in watermelon

As a prerequisite for sexual reproduction,gametophyte development is an interesting process involving cell proliferation,differentiation and specialization.Gametogenesis has been extensively explored in model plants,but the regulatory mechanism of gametophyte development largely remains unknown in Cucurbitaceae species.In present study,we have shown that watermelon ClBBM and ClPLT2,two AP2/ERF transcription factors,participated in both male and female gametophyte development.Clbbm and Clplt2 single mutants resembled wild-type phenotypes in both vegetative and reproductive development.But Clbbm/Clplt2 double mutant showed partial pollen abortion and bore less seeds comparing to WT.Our results indicated that the abnormal pollen grains were caused by premature tapetum degeneration,and reduced seed-set was due to faulty embryo sac development.ClBBM and ClPLT2 were expressed in FG4 embryo sacs,and their transcripts were also detectable in the tapetum and microspore of stage 9 anthers,which was consistent with developmental stages of defective phenotypes observed in double mutant.The expression of genes essential for tapetum development,ClATM1,ClAMS,ClMS1 and ClMS188,was decreased in Clbbm/Clplt2 double mutants.Moreover,the transcriptome analysis indicated that ClBBM and ClPLT2 participated in tapetum and pollen wall development by regulate cell cycle,transmembrane transport,glucan and cellulose metabolic process.Collectively,ClBBM and ClPLT2 were functionally redundant in regulating gametophyte development in watermelon,and their functions differ from their homologous genes in model plant Arabidopsis.

3种生长调节剂对克瑞森葡萄硬枝扦插的影响

【目的】探索不同生长调节剂及其浓度对克瑞森葡萄硬枝扦插成苗的影响,以期为克瑞森葡萄苗木扦插繁育提供适宜的生长调节剂种类及其浓度。【方法】以克瑞森葡萄1年生硬枝为试材,采用不同浓度的萘乙酸(NAA)、吲哚乙酸(IAA)和吲哚丁酸(IBA)溶液处理进行扦插试验,以清水处理为对照,测定不同处理对克瑞森葡萄插条生根及萌芽生长的影响。【结果】3种不同浓度的植物生长调节剂对克瑞森葡萄插条的萌芽与生根均有显著促进作用。3种生长调节剂处理插条萌芽率都很高,2000 mg·L^(-1)NAA和2000 mg·L^(-1)IAA处理过的插条生根率较好,但500 mg·L^(-1)IBA处理的插条单株根数最多,根长较长。【结论】试验中,500mg·L^(-1)IBA处理对克瑞森葡萄硬枝扦插效果最好。

Influence of Gamma Irradiation on Seedless Citrus Production: Pollen Germination and Fruit Quality

Seedlessness is a desirable characteristic in citrus fruits sold for fresh consumption. Gamma irradiation is widely used to obtain seedless citrus fruits. Here, different clones of the self-incompatible parthenocarpic ‘Moncada’ mandarin, obtained by gamma irradiation, were studied to assess seedlessness, pollen germination, fruit characteristics and qua- lity attributes. Findings indicate that irradiation altered aspects other than seedlessness, such as pollen germination, and some of the clones presented different weight, size, acidity and maturity index. Fruit quality and nutritional bio- components were affected differently;some clones presented no changes compared to the control ‘Moncada’ man-darin, while other clones showed significant differences. In general, all clones examined presented low seed numbers and re- duced pollen viability. Some of these clones, which ripen late in the season and whose fruit quality is maintained or improved, are in the process of registration.

Influence of Water Stress on the Citric Acid Metabolism Related Gene Expression in the Ponkan Fruits

To explore the influence of water stress on fruit quality and gene expression related to citrate metabolism of ponkan. The test were conducted from May 15 to December 24 in 2013 using six-year-old ponkan (C. blanco cv. Ponkan) trees with 40% soil water conditions by taken regular watering as control. The content of acids in fruit were determined by HPLC, and relative expression of related genes of citric acid metabolic were determined by relative fluorescence quantitative PCR. The results showed that the content of citric acid, malic acid, quinic acid and total organic acids per gram sarcocarp were extremely increased by 285.2%, 320%, 480% and 299.1%, and the content of per-fruit organic acid were 77.39%, 89.64%, 117.24% and 75.9% respectively compared to those control in the fruit mature stage. Relative expression of CitCS1, CitCS2 were higher than control, and relative expression of CitAco1, CitAco2, CitAco3 had a certain increase in the late fruit development, were lower in mature stage. Three relative expression of CitIDH gene were higher than control in mature stage. Low CitGAD4 relative expression and undetectable in mature stage, the relative expression of CitGAD5 gene had a role in promoting under water stress. Furthermore, the relative expression of CitCS1, CitCS2, CitACO1, CitACO3, CitIDH1, CitIDH2, CitIDH3, CitGAD4 and CitGAD5 were influenced by water stress through the correlation analysis. Water stress caused the accumulation of citric acid, declined fruit quality, leaded to change of the genetic rela- tive expression about citric acid synthesis and degradation. The down-regulation of CitACO1, CitGAD4 and up-regulation of CitCS1, CitCS2 might be one of the reasons that promoted to the accumulation of citric acid.

‘Fresno Seedless’葡萄幼胚和胚乳发育及败育的组织学观察

以种子败育型无核葡萄品种‘Fresno Seedless’为试材,采用石蜡切片技术和铁矾媒苏木精染色法,对‘Fresno Seedless’葡萄的幼胚和胚乳的发育及败育过程进行了系统的细胞学观察,以期确定其幼胚和胚乳开始败育的时期。结果表明:‘Fresno Seedless’葡萄合子于花后16d开始分裂,经过二细胞原胚、多细胞原胚,发育至球形胚时期,开始退化败育,时间为花后38d;‘Fresno Seedless’葡萄初生胚乳核于花后6d开始分裂,待胚乳细胞基本充满胚囊后开始退化败育,时间为花后32d。胚囊内胚乳首先从胚囊珠孔端开始败育,随后合点端胚乳开始败育。因此,‘Fresno Seedless’葡萄的胚挽救最佳取样时期是花后32～36d。该研究结果为利用‘Fresno Seedless’葡萄做母本进行胚挽救育种的大田取样时期提供了较为科学的参考依据。

Rapid Seedless Synthesis of Gold Nanoplates with Microscaled Edge Length in a High Yield and Their Application in SERS

We report a facile and reproducible approach toward rapid seedless synthesis of single crystalline gold nanoplates with edge length on the order of microns.The reaction is carried out by reducing gold ions with ascorbic acid in the presence of cetyltrimethylammonium bromide(CTAB).Reaction temperature and molar ratio of CTAB/Au are critical for the formation of gold nanoplates in a high yield,which are,respectively,optimized to be 85 °C and 6.The highest yield that can be achieved is 60 % at the optimized condition.The synthesis to achieve the microscaled gold nanoplates can be finished in less than 1 h under proper reaction conditions.Therefore,the reported synthesis approach is a time-and costeffective one.The gold nanoplates were further employed as the surface-enhanced Raman scattering substrates and investigated individually.Interestingly,only those adsorbed with gold nanoparticles exhibit pronounced Raman signals of probe molecules,where a maximum enhancement factor of 1.7 9 10~7 was obtained.The obtained Raman enhancement can be ascribed to the plasmon coupling between the gold nanoplate and the nanoparticle adsorbed onto it.

QTL-seq analysis of the seed size trait in grape provides new molecular insights on seedlessness

Seedlessness in grape(Vitis vinifera)is an important commercial trait for both the fresh and drying markets.However,despite numerous studies,the mechanisms and key genes regulating grape seedlessness are mostly unknown.In this study,we sequenced the genomes of the V.vinifera seeded cultivar‘Red Globe’,the seedless cultivar‘Centennial Seedless’,and the derived hybrids.Nonsynonymous single nucleotide polymorphisms(SNPs)were identified by genome sequencing and analyzed using published transcriptome data.Nonsynonymous SNPs occurred in genes related to seed development,which were identified as protein kinases,transcription factors,and cytochrome P450 s and showed differential expression during ovule development in both seeded and seedless grapes.These nonsynonymous SNP-associated genes were mainly involved in biological processes such as hormone balance,seed coat and endosperm development,reproductive organ development,oxidation and reduction,senescence and cell death.A potential quantitative trait locus(QTL)region associated with seed size was characterized based on the SNP-index,and expression analysis of candidate genes in the QTL region during ovule development in multiple seeded and seedless grape cultivars were conducted.Three SNPs were further subjected to SNa Pshot analysis and one SNP in G8 showed 67.5%efficiency in the grape progeny validation.Overall,the data obtained in this study shed light on the differences in seed development between seeded and seedless progeny at the genomic level,which provides valuable resources for future functional studies and grape breeding.

Development and application of KASP marker for high throughput detection of the seedless trait in grapevine

Molecular marker-assisted selection(MAS) can significantly accelerate and improve the efficiency of the breeding process in seedless grape cultivars. In this study, we developed the KASP_VviAGL11 and VviAGL11_410 markers based on a single nucleotide polymorphism(SNP) site(Chr18: 26889437(A/C)) of the VviAGL11 gene, and compared them with previously reported SSR markers p3_VvAGL11 and 5U_VviAGL11 by testing 101 cultivars and 81 F1 hybrid progenies. The results showed that both of the proposed markers obtained 100% accuracy rates in detecting allele A, which was closely associated with the seedless trait in grapes, while p3_VvAGL11 and 5U_VviAGL11 had lower accuracy rates due to their tendency to produce false positives. After careful evaluation of the technical advantages and disadvantages associated with these markers, we concluded that KASP_VviAGL11 was superior in terms of simplicity,cost-effectiveness, efficiency, and accuracy. Thus, we optimized the process of molecular MAS for seedless grapes,focusing on the KASP_VviAGL11 marker as a central component, to provide key technical support for the development of new seedless grape cultivars.

Cloning and Sequence Analysis of a Cysteine Proteinase Inhibitor Gene from Seedless Litchi

[Objective] This study aimed to clone and analyze the cysteine proteinase inhibitor gene from seedless litchi. [Method] According to the EST se- quence of cysteine proteinase inhibitor in constructed SSH snhtraetive library of seedless litchi abortion, nucleotide sequence of the cysteine proteinase inhibitor gene was obtained by using RACE technology and analyzed by using bioinformatics software. [ Result ] A cysteine protease inhibitor gene was obtained with the sequence of 635 bp containing a 321 bp open reading frame. It was predicted that the erlcoded protein contained 106 amino acids with conserved domain of cysteine proteinase inhibitor and had relatively high homology with the cysteine proteinase inhibitor gene of several species, [ Conclusion] This study laid the foundation for further ex- ploring the physiological functions of this cysteine proteinase inhibitor gene in plants.

Immature Embryo Rescue and in Vitro Development Evaluation of Intraspecific Hybrids from Brazilian Seedless Grapevine“Superior×Thompson”Clones

The fruit production for export is an economically significant activity in the Valley of S?o Francisco River, especially in the irrigated lands of Petrolina-PE/Juazeiro-BA, Brazil. The development of new genetic material most suitable to the tropical climate and the demands of the consumer market have led to the selection of new seedless grapes cultivars. In this case, the use of the embryo rescue technique has produced satisfactory results for obtaining such materials, especially in the semiarid region. This study aimed to evaluate the in vitro development of intraspecific hybrids of grapevine (Vitis vinifera L.), derived from the rescue of immature embryos resultant from the crossing of “Superior Seedless” and “Thompson Seedless” Brazilian clones. To establish and develop the cultivation, the culture media was supplemented with 30 g/L sucrose, 0.1 g/L myo-inositol, 0.002 g/L glycine, 0.1 mg/L indoleacetic acid (IAA), 6.5 g/L of agar, adjusted pH to 5.7. The experiment was evaluated after 90 days. The variables measured were: number of nodes, number of leaves, plant height (cm), number of roots and length (cm) of the root system and internodes. The period of 60 days of in vitro culture of ovules resulted in the highest values of embryos (about 50%), as well as better characterized developmental stages with higher germination (47.3%). The three types of hybrid grapes evaluated in micropropagation showed very similar values of the measured parameters, even having originated from embryos of different developmental stages.

Effect of Different Shading Practices Used at Pre-harvest Stage on Quality and Storage Life of Sultana Seedless Grapes

Covering the grapevine rows to delay maturity and thus the harvest date becomes a widespread practice at Sultana seedless vineyards. The research work was carried out with different covers to examine their effects on grape quality and storage life in 2009 and 2010. Grapevines were covered with three different densities （shading ratios： 35%, 55%, 75%） at the veraison period. Unshaded （0%） control grapevines and grapevines shaded under different ratios were covered with polypropylene cross-stitch just before harvest maturity. Grapes under cover were harvested nearly one month after than those maturing under open conditions. Grape clusters were packed in PE bags and pre-cooled after harvest and then SO2-generating pads has been put on top, bags were sealed. In the second year at the 120th day of the storage, decay development has been observed in all applications. Total soluble solids content, maturity index and a＊ colour value were found lower in 55% and 75% shaded grapes compared to unshaded grapevines. Still, berry removal force and hue angle value in 55% and 75% of shaded grapes were higher than unshaded grapes. Effects of shading ratios on these parameters decrease or disappear at the end of storage.

留树贮藏对椪柑果实品质和呼吸强度的影响

【目的】探明椪柑留树贮藏果实的品质变化特征,为延长椪柑鲜果供应期提供理论依据。【方法】以留树贮藏椪柑果实为研究对象,以室温贮藏果实为对照,利用液相色谱技术、气相色谱和色度计等果实品质检测方法,研究留树贮藏方式对椪柑果实可溶性固形物、果皮色泽、总糖、有机酸和异味等品质和呼吸强度的影响。【结果】椪柑果实留树贮藏40~80 d可溶性固形物含量高于对照,其中60和70 d时分别显著高于对照14.06%和17.28%;总糖在20、40、50和70 d分别比对照低23.31%、6.24%、17.65%和18.82%;留树贮藏10~80 d时柠檬酸含量分别是对照的1.26~1.92倍;红色饱和度a^(*)值降低(30~80 d)2.13~5.03,果皮颜色偏黄,呈橘黄色;除贮藏40 d外,留树贮藏呼吸强度均较对照高1.16%~127.54%;贮藏80 d时异味的代表性物质乙醇含量达峰值(293.28μg/g),室温贮藏是留树贮藏的3.29倍。【结论】生产中建议留树贮藏最佳时间在1月中下旬至2月初春节前后,留树贮藏椪柑果实能够保持色泽橘红,固酸比适宜,风味浓郁,效益好。

Effects of Different Concentrations of Paclobutrazol on Seedling Quality of Small Seedless Watermelon in Autumn

[Objectives]The paper was to explore the effects of different concentrations of paclobutrazol on seedling quality of small seedless watermelon in autumn.[Methods]The leaves of watermelon seedlings were sprayed with 30,50,80 and 120 mg/L paclobutrazol at cotyledon expansion stage,and those sprayed with tap water were used as control.[Results]All the treatments of paclobutrazol effectively inhibited the excessive growth of watermelon at seedling stage,significantly enhanced the leaf quality of watermelon seedlings and promoted root growth.The seedlings treated by 50 mg/L paclobutrazol had the best performance compared with the control:a decrease of hypocotyl by 37.9%,an increase of plant dry weight by 46.55%,significantly increased seedling strength index,deep green leaf color,thick leaves,an increase of specific leaf weight by 28.13%,and significantly heavier root weight and longer length.[Conclusions]The results will provide a reference for strong seedling cultivation of small seedless watermelon.

Optimization of Molecular Distillation for Removal of Limonene from Ponkan Essential Oil Using Response Surface Methodology

Ponkan (Citrus reticulate Blanco) is one of the most popular citrus fruits for daily consumption and processing in China. In this study, limonene was removed from ponkan essential oil (PEO) by using an effective methodology to estimate the optimum levels of three independent variables of molecular distillation (MD) in order to minimize the limonene content and (b) maximize the removal rate. A three-factor Box-Behnken design (BBD) was employed to evaluate the effect of distilling temperature (45.00~55.00 ℃), wiper rolling speed (150.00~250.00 r/ min) and distilling pressure (25.00~35.00 Pa). The optimum parameters within the experimental range of the variables were extrapolated to be: distilling temperature 47.00℃, wiper rolling speed 203.00 r/min and distilling pressure 30.00 Pa. Optimal values extrapolated by response surface methodology (RSM) for the limonene content and removal rate of the desirable PEO were 47.82% and 84.67%, respectively.

Influence of Bioregulator Gibberellic Acid on Some Technological Characteristics of Cluster and Berry from Some Seedless Grape Varieties

Solutions of gibberellic acid prepared at three concentration levels including 5, 10 and 20 mg/L, were applied at two seedless grape varieties, Thompson and Belgrade, by spraying, during the three different periods of the vine growing： before blooming, after blooming and before veraison, in order to study their influence on some cultural technological characteristics. Dimension and shape of the cluster and berry, mechanical characteristics of the berries and chemical content of the must （sugar and total acids） has been also investigated. It was noticed that the concentration of gibberellic acid had influence on the technological characteristics of the berries in all grape growing periods tested. The addition of gibberellic acid at concentration of 20 mg/L increased the weight of the cluster and berry, and increased the transportability of the berries belonging to the two seedless varieties.

Screening Test of Small Seedless Watermelon Varieties in Beijing Area

[Objectives]This study was conducted to select small seedless watermelon varieties with excellent quality and high yield suitable for planting in Beijing.[Methods]Under the conditions of facility cultivation in Beijing,a screening test was conducted on seven small seedless watermelon varieties for growth process,field traits,yield traits,fruit quality,and disease resistance.[Results]Varieties Mitong,Motong and Tianbao Xiaowuzi had a short growth period and relatively stable growth,and bore fruit easily,and their female flowers were closely spaced.Their yields exceeded 60000 kg/hm~2,ranking in the top three among all varieties.The fruit had a thin skin,a bright red flesh color,and a high mass fraction of soluble solids in the center.The flesh was tight or crispy,and tasted good.They also showed good disease resistance.Overall,they showed excellent comprehensive agronomic traits,and are thus suitable for facility cultivation in Beijing and its surrounding areas.[Conclusions]This study provides a theoretical basis for the upgrading of varieties and the increase in production and farmers'income in the suburbs of Beijing.