Induction and Genetic Identification of Embryogenic Calli from Hybrids of Shatian Pummelo

Shatian pummelo （Citrus grandis L. Osbeck cv. Shatian） is an elite variety in China, and the regeneration of the embryogenic callus is difficult. Diploid Shatian pummelo was used as the female and crossed with the allotetraploid somatic hybrid NS （Nova Tangelo ＋ Succari Sweet orange）, [ （ C reticulata Blanco x C. paradisi Macf.） cv. Nova ＋ C sinensis L. Osbeck cv. Succari]. About 90 days after pollination, the embryos obtained from crosses were cultured on the solid media of MT ＋ ME （malt extraction, 500 mg L^-1） and MT ＋ GA3 （1 mg L^-1）. The embryogenic callus was initiated from the embryoids and plantlets＇ hypocotyls and could be subcultured. Flow cytometry and SSR analysis verified that the callus was from the triploid hybrids. The callus had embryogenesis capacity and produced a large number of embryoids on MT ＋Lactose （50 g L^-1） medium after being subcultured for two years. It is comparatively easier to obtain the callus from the hybrid embryo than from Shatian pummelo itself. The callus is valuable for the conservation and utilization of Shatian pummelo.

Assessment of the Genetic Diversity of Pummelo Germplasms Using AFLP and SSR Markers

The genetic diversities of 110 pummelo germplasms and 12 of their relatives were analyzed by SSR and AFLP methods. Approximately 99.1% of the 335 SSR loci were polymorphic, and 9.85 alleles per SSR locus were identified. The gene diversity values changed from 0.1939 to 0.9073, and 46 SSR polymorphic bands were scored. 72% of the 343 AFLP loci were polymorphic, and 82 polymorphic loci per AFLP were identified. Heterozygosity changed from 0.21863 to 0.28445, and 44 AFLP polymorphic bands were scored. The UPGMA result showed that 122 pummelo genotypes and their relatives could be divided into eight groups, and the pummelo genotypes composed mainly of Shatian pummelo varieties group, Wendan pummelo vareties group and a huge hybrid pummelo varieties group. The classification result was expected to widen the genetic background of pummelos using various target varieties.

Quality and Nutrition of Pummelo as Influenced by Potassium

This study aimed to determine the influence of different rates of K （potassium） and K foliar application on the quality and plant nutrition condition of 13-year old ＇Magallanes＇ pummelo trees [Citrus maxima （Burm. ex Rumph.） Merr.]. The experiment was conducted at SODACO （South Davao Corporation） farm, Davao City, Philippines from March to October 2010. Five treatments with increasing K levels were applied per tree： control, no K, 150 g K basal, 225 g K basal and 225 g K basal ＋ foliar application. Application of 225 g K rates positively influenced fruit quality of pummelo. Fruit pulp diameter and juice weight increased while peel thickness decreased. The 225 g K rates also increased juice pH, TA （titrable acidity） and TSS （total soluble solids） per tree. The yield of total phenol, flavonoid, vitamin C and oil per tree increased with 225 g K application. Results of the soil analysis before and after the conduct of the study showed an evidence of fertilizer absorption by the trees. It was also observed that foliar fertilization reduced soil acidity. This indicates the important role of K in improving the nutrient utilization and quality of ＇Magallanes＇ pummelo.

Preliminary Study on Quality Standard of Pummelo Pericarp from Guangxi

[Objectives]This paper aimed to study the quality standard of pummelo pericarp from Guangxi.[Methods]The medicinal material was identified by microscopy and thin-layer chromatography,and the contents of moisture,total ash,acid-insoluble ash and extract in the medicinal material were determined according to the methods of the Chinese Pharmacopoeia(2010 edition).[Results]The content limits for moisture,total ash,acid-insoluble ash and extract were determined(moisture≤12%;total ash≤6.0%;acid-insoluble ash≤0.6%;water-soluble extract≥43.0%;naringin≥10.00 mg/g).The micro-characteristics of pummelo pericarp were identified,and a thin-layer chromatography identification and content determination method was established.[Conclusions]The established method can provide an experimental basis for the formulation of quality standard for pummelo pericarp.

Phytochemical Composition,Antioxidant and Antibacterial Properties of Pummelo(Citrus maxima(Burm.))Merr.against Escherichia coli and Salmonella typhimurium

The antioxidant and antibacterial activities of the phytochemical constituents of the pericarp, mesocarp and segment membrane crude ethanolic extracts of Pummelo (Citrus maxima (Burm.)) fruit were were tested against Escherichia coli and Salmonella typhimurium. Preliminary phytochemical test revealed the presence of phenols, tannins, saponins expressed as catechine quivalent (CE)/100ml and flavonoid expressed as gallic acid equivalent (GAE)/100ml. The order of which was as follows pericarp > segment membrane > mesocarp. The strongest antioxidant activity was obtained by the pericarp extract (29.64 expressed as % lipid peroxidation). The differences in the measured amount of phytochemicals and antioxidant activity among the three sample extracts were found to be significant. In terms of antimicrobial activity, the pericarp, mesocarp and segment membrane extracts generated zone of inhibitions measuring 17.10, 18.00 and 17.03 mm for S. typhimurium, respectively at 100% concentration. E. coli was noted to be inactive in all three sample extracts at 100% concentration. The capacity of E. coli to counteract the inhibitory effect of the phytochemicals contained in the pummelo extracts may be attributed to its rough corrugated cell wall and thick periplasmic space as opposed to the smooth curved and barely seen periplasmic space of S. typhimurium. However, no significant correlation was detected among the phytochemical content, antioxidant and in vitro antimicrobial activities of the sample extracts against S. typhimurium.

基于Spring MVC的化橘红病虫害防治技术数据库系统的设计

文章设计了化橘红病虫害防治技术数据库系统,并详细分析了设计流程。系统界面设计友好,操作便捷,收集了与化橘红相关的病虫害信息及防治方法,能够为果农提供参考,具有一定的应用价值。

柚和葡萄柚花粉直感对东试早柚果实生长及品质的影响

【目的】针对东试早柚果实个体差异大、品质不稳定等问题,通过异花授粉探究花粉直感对东试早柚果实品质的影响,并对各项指标进行综合评价,旨在筛选出能够有效改善东试早柚果实品质的授粉品种,为生产上授粉品种配置和品质提升提供理论依据。【方法】以23年生东试早柚为母本,柚和葡萄柚不同品种为父本,进行人工授粉,比较不同授粉组合的坐果率、果实生长、果实品质差异,并采用隶属函数法进行综合评价。【结果】通过比较分析发现果实横径和纵径生长动态呈“S”形;异花授粉显著提高东试早柚坐果率,坐果率均高于60%。在果实外在品质方面,Flame授粉对果实单果质量提升效果不明显,但其他授粉组合果实单果质量比自然授粉显著提升27.77%~85.08%;在所有授粉组合中,自然授粉果实横径和纵径最小,Sarawak授粉果实横径和纵径显著高于自然授粉和其余品种授粉果实。异花授粉能够降低果实裂瓣数,增加种子数,且不同品种授粉种子直感存在差异。在果实内在品质方面,Oroblanco授粉能提高可溶性固形物含量,但Sarawak、Tahition、泰国红宝石柚、越南青柚授粉显著降低果实可溶性固形物含量,显著提高总酸含量;不同授粉组合间果实可溶性蛋白含量无显著差异,但可溶性糖、维生素C以及木质素、柚皮苷含量存在差异,表现出明显花粉直感效应。果实种子数与果实横径呈显著正相关,裂瓣数与果实横径和种子数呈显著负相关;外在品质与内在品质主要指标间呈显著负相关。通过主成分分析法和隶属函数法综合评价结果显示,Genetic Dwarf、Oroblanco授粉果实综合得分为0.68、0.67,高于自然授粉0.66。【结论】异花授粉能够提高坐果率,同时对果实生长发育和果实品质有显著影响。Genetic Dwarf和Oroblanco授粉在一定程度上能够改善东试早柚果实综合品质,可为生产上授粉品种配置、提高果实品质提供参考依据。

以雄性不育华柚2号为母本有性杂交创制柚潜在无核新种质

【目的】中国柚地方品种多但多数有种子,缺乏品质优良且无核的柚新品种。柑橘胞质雄性不育为母系遗传,以具有该性状的柚为母本与有核柚杂交,可将雄性不育胞质遗传至有性后代,有望在二倍体水平实现柚无核新品种选育。【方法】以利用细胞工程技术自主创制的雄性不育胞质杂种柚华柚2号为母本、地方特色有核柚为父本进行有性杂交,果实成熟期收集种子后,催芽播种培育杂交后代;在幼苗阶段依据多倍体形态特征从后代中筛选疑似多倍体并用流式细胞仪对其倍性进行鉴定;通过亲本重测序开发多态性InDel标记并对杂交后代遗传来源进行鉴定。【结果】以华柚2号为母本,贡水白柚、雷公柚、菊花芯柚、马家柚和甜柚1号为父本,配置5个杂交组合;人工授粉后共获得65个成熟果实,收集到1163粒种子,通过催芽播种共获得实生苗781株;依据多倍体形态特征,从华柚2号×甜柚1号杂交组合的110株后代中发掘出2株疑似多倍体,经流式细胞仪倍性鉴定其中1株为四倍体。用6对多态性InDel引物对从华柚2号×贡水白柚和华柚2号×雷公柚随机选取的各44株实生后代进行遗传鉴定,结果表明,其均有各自父本的特异条带,暗示2个杂交组合所有后代均为其相应双亲的有性后代。【结论】以雄性不育胞质杂种柚为母本创制的有性后代,为二倍体水平的柚无核新品种选育提供了宝贵的种质材料。

琯溪蜜柚疏果黄酮酶法辅助提取工艺优化及其抑制酪氨酸酶活性

本论文旨在探讨β-葡萄糖苷酶法辅助提取琯溪蜜柚疏果黄酮的工艺及提取得到的黄酮对酪氨酸酶抑制作用。以总黄酮提取量为指标,采用单因素实验及响应面试验对β-葡萄糖苷酶酶活力、酶解pH、温度及时间进行优化,并研究总黄酮提取物对酪氨酸酶的抑制作用。结果表明,通过单因素实验和响应面试验优化得到琯溪蜜柚疏果黄酮提取的最优条件是:β-葡萄糖苷酶酶活力0.0128 U/mL、酶解pH4.4、酶解温度35℃、酶解时间4.8 h,在该工艺条件下疏果总黄酮提取量为34.0 mg/g;该黄酮提取物对酪氨酸酶的半抑制浓度(IC50)为0.31 mg/mL。本研究结果说明酶法工艺可显著提高琯溪蜜柚疏果黄酮的提取量,且琯溪蜜柚疏果黄酮具有较强的酪氨酸酶抑制作用,研究结果为琯溪蜜柚疏果黄酮提取及应用提供了参考依据,有利于促进琯溪蜜柚的高值化利用。

重构本草——化橘红

通过古籍文献调研、中医药现代研究成果的梳理,结合中医临床及中药学等学科领域内知名专家学者的认识及应用经验,归纳出化橘红:功效主要为化痰止咳、理气宽中。症靶为咳嗽。现代药理研究发现化橘红及其有效成分具有化痰止咳、抗呼吸道炎症、降糖调脂、抗阿尔茨海默病、防治糖尿病心肌功能损伤等作用。化橘红基本无毒副作用,阴虚燥咳及久嗽气虚者应慎用。临床医家常用剂量为10~30 g,内服时当根据不同疾病,或同一疾病的不同阶段进行辨证使用,并积极探索化橘红用量与症靶、标靶的量效关系构建。

化橘红活性成分及功效研究进展

化橘红为芸香科植物化州柚(Citrus grandis'Tomentosa')或柚(Citrus grandis(L.)Osbeck)未成熟或近成熟的干燥外层果皮,因其药效独特而被誉为“十大广药”之一。化橘红中含有大量的黄酮、多糖、挥发油等活性物质,具有明显的抗氧化、抗炎、保肝护肾、降脂减糖、调节免疫等功效。对化橘红活性成分、提取方法及其功效等进行了综述,以期为化橘红在保健食品、医药等领域的应用提供参考。

A novel histone methyltransferase gene Cg SDG40 positively regulates carotenoid biosynthesis during citrus fruit ripening

The flesh color of pummelo(Citrus maxima)fruits is highly diverse and largely depends on the level of carotenoids,which are beneficial to human health.It is vital to investigate the regulatory network of carotenoid biosynthesis to improve the carotenoid content in pummelo.However,the molecular mechanism underlying carotenoid accumulation in pummelo is not fully understood.In this study,we identified a novel histone methyltransferase gene,CgSDG40,involved in carotenoid regulation by analyzing the flesh transcriptome of typical white-fleshed pummelo,red-fleshed pummelo and extreme-colored F1 hybrids from a segregated pummelo population.Expression of CgSDG40 corresponded to flesh color change and was highly coexpressed with CgPSY1.Interestingly,CgSDG40 and CgPSY1 are located physically adjacent to each other on the chromosome in opposite directions,sharing a partially overlapping promoter region.Subcellular localization analysis indicated that CgSDG40 localizes to the nucleus.Overexpression of CgSDG40 significantly increased the total carotenoid content in citrus calli relative to that in wild type.In addition,expression of CgPSY1 was significantly activated in overexpression lines relative to wild type.Taken together,our findings reveal a novel histone methyltransferase regulator,CgSDG40,involved in the regulation of carotenoid biosynthesis in citrus and provide new strategies for molecular design breeding and genetic improvement of fruit color and nutritional quality.

‘红肉蜜柚’果实汁胞粒化过程中POD同工酶谱分析

【目的】探讨‘红肉蜜柚’果实汁胞粒化过程中过氧化物酶(POD)的同工酶变化及赤霉素对其的影响。【方法】以赤霉素处理和未经赤霉素处理的‘红肉蜜柚’果实的近中柱和远中柱汁胞为材料,统计粒化指数后,采用垂直板聚丙烯酰胺凝胶电泳方法进行可溶性和离子结合POD同工酶谱测定。【结果】发现‘红肉蜜柚’果实汁胞粒化指数随着果实生长发育呈先平稳后急剧上升的趋势,有无赤霉素处理无明显变化;可溶性POD中存在酸性POD和碱性POD,有无赤霉素处理对近、远中柱POD出现的时期和组分基本没有影响;离子结合POD只存在碱性POD,近中柱POD同工酶谱分离出6条(5条为果实发育后期的特异性谱带)、远中柱POD分离出4条,两者出现的时期和组分差异较大,赤霉素处理使近中柱5条果实发育后期特异性谱带减为3条且时期提前,而赤霉素处理下的远中柱没有检测到谱带。【结论】粒化时期,离子结合POD出现的5个特异性碱性POD可能与汁胞粒化关系密切,赤霉素处理可明显减少POD谱带,但赤霉素对汁胞粒化的影响有待进一步研究。

早熟优质蜜柚新品种——红肉蜜柚的选育

红肉蜜柚系琯溪蜜柚优变芽系选育而成。果大、无子、空心、皮薄;风味酸甜适口,肉质细嫩化渣,总糖、总酸、维生素C含量分别为8.76%、0.74%、0.38mg/g;汁胞呈红色,色素成分主要为β-胡萝卜素与番茄红素,含量分别达(72.15±17.13)、(41.10±2.24)μg/g,叶黄素微量;成熟期9月下旬至10月初;盛产树666.7m2产量2000kg以上。

‘红肉蜜柚’果实汁胞粒化与不同组分POD活性的关系

为了研究‘红肉蜜柚’[Citrus grandis(L.)Osbeck.cv.Hongrou miyou]果实汁胞粒化过程中不同结合态过氧化物酶(POD)的变化,以及赤霉素处理对此的影响。以赤霉素处理和未经赤霉素处理的‘红肉蜜柚’果实的近中柱和远中柱汁胞为材料,统计粒化指数后,进行POD提取体系筛选与酶活测定。发现‘红肉蜜柚’果实汁胞粒化指数逐渐上升,有无赤霉素处理无明显变化;可溶性POD最佳提取体系为pH8.0,0.1 mol/L Tris-HCl的缓冲液;离子结合POD的为含0.1%Triton和1 mol/L Na Cl的pH8.0,0.1 mol/L Tris-HCl缓冲液;未检测到共价结合POD活性。此外,可溶性、离子结合POD活性在生长发育过程中整体呈下降趋势,且无论是否经赤霉素处理,近中柱与远中柱汁胞的可溶性POD活性均存在极显著差异(p<0.01),而离子结合POD活性均无显著差异(p>0.05);无论近中柱或远中柱,有无赤霉素处理的可溶性POD均存在极显著差异(p<0.01),而离子结合POD活性仅在远中柱汁胞中有极显著差异(p<0.01)。所以,可溶性POD可能与汁胞粒化关系密切,而离子结合、共价结合POD和赤霉素对汁胞粒化的影响有待进一步研究。

柚皮中柚皮苷的乙醇提取工艺研究

目的 探讨柚皮中柚皮苷的乙醇提取最佳工艺条件。方法 运用正交试验法考察乙醇浓度、提取温度、提取时间和乙醇用量对提取率的影响 ,采用两步结晶法对提取物中的柚皮苷进行精制。结果 柚皮中柚皮苷的乙醇提取最佳工艺条件为 2 5倍原料重的 70 %乙醇、6 0℃条件下保温浸提 1h;两步结晶法所得柚皮苷精制品的纯度为90 .0 1%。结论 采用本实验所确定的提取及精制方法可使柚皮中柚皮苷得到有效提取和分离。

柚果皮中生理活性成分研究进展

对柚果皮中黄酮类化合物、类柠檬苦素、天然色素及香精油的化学组成、分布。

沙田柚杂交后代群体的SSR鉴定与遗传多样性分析

【目的】利用SSR标记对沙田柚杂交后代群体进行杂种鉴定和遗传多样性研究,为柑橘种质创新和遗传改良提供技术、材料和理论支持。【方法】利用SSR技术分析沙田柚两个杂交组合159株后代的杂种性质,采用UPGMA聚类分析法分析后代群体和亲本的遗传关系。【结果】4对引物可以从104株沙田柚×强德勒柚的杂交后代鉴定出103株真杂种,鉴定率达到99.04%,且在引物AGC9的扩增图谱中有70个单株出现了双亲都没有的新条带;5对引物可鉴定出沙田柚×红江橙的全部杂交后代均为真杂种,一个纯合显性标记AAT12被发现,在引物GA18和AGC9的扩增结果中出现了亲本位点的缺失;UPGMA聚类分析结果显示,两个杂交组合后代群体均出现较显著的遗传变异,两个组合的遗传多样性也有较大的差异。【结论】SSR标记适合柑橘杂种的鉴定和遗传多样性分析,杂交是获得柑橘遗传变异株系的有效途径。

十种柚类及柚杂种果实中类黄酮含量的超高效液相色谱分析

【目的】利用超高效液相色谱法同时检测10种柚及柚杂种果实中11种类黄酮含量,测定分析各品种果实类黄酮的种类及其含量变化规律。【方法】超高效液相色谱法,色谱柱:ACQUITY UPLC BEH C18分析柱(2.1 mm×100 mm,1.7μm);流动相:0.1%乙酸水溶液(A)和甲醇(B),采用梯度洗脱,洗脱程序为:(1)0—5 min,90%—80%A;(2)5—10 min,80%—30%A;(3)10—15 min,30%—20%A;(4)15—16 min,20%—90%A;柱温:35℃;流速:0.3 mL.min-1。【结果】类黄酮组分及含量与柑橘的遗传基因型有关,柚果实中类黄酮以柚皮苷为主,葡萄柚果实中以柚皮苷和新橙皮苷为主。柚皮苷在瑞红中含量最高(果皮:27.5 mg.g-1DW,果肉:2.73 mg.g-1DW),新橙皮苷在鸡尾中含量最高(果皮:14.93 mg.g-1DW,果肉:0.70 mg.g-1DW)。杂种后代类黄酮累积受其亲本影响。【结论】超高效液相色谱法是一种定性定量柑橘果实类黄酮的有效方法,柚果实中类黄酮含量丰富且各种类之间组分含量差异显著。

中国柚类生态分布多样性研究

利用分子标记AFLP技术分析了我国柚类生态分布遗传多样性,根据我国自然形成的西南及长江流域、东南沿海、华南柚产区等3个分布带,其中种群内平均预期杂合度为0.210 4,种群间平均预期杂合度为0.006 8,群体内遗传分化程度Fst为0.031 1,即种群内的遗传多样性所占的比例为96.87%,种群间变异占总变异的3.13%,3个中心产区间基因流Nm为7.79。以西南及长江流域遗传多样性最高,其次为东南沿海,华南柚产区遗传多样性最小。按照内陆性与海洋性两大系统进行柚类遗传多样性分析,所有位点总的预期杂合度为0.224 7,其中种群内平均预期杂合度为0.219 1,种群间平均预期杂合度为0.005 7,群体内遗传分化程度Fst为0.025 1,即种群内的遗传多样性所占的比例为97.51%,种群间变异占总变异的2.5%,2个系统间基因流Nm为9.7。预期杂合度内陆性系统(0.229)高于海洋性系统(0.209)。