Effects of Lanthanum and Europium on Rooting of Plantlet Eriobotrya Japonica Lindl. in vitro

The effect of La 3+ and Eu 3+ on the rooting of Eriobotrya japonica Lindl. plantlet in vitro was studied with adding La 3+ and Eu 3+ to the rooted medium. The rooting rate, the number of root and the length of root were studied after transplanting 20 d. The activity of peroxidase, nitrate reductase and fresh weight of roots were determined after transplanting 44 d. The results show that the optimum concentration range of La 3+ (1.0～3.0 μmol·L -1), Eu 3+ (2.0～3.0 μmol·L -1) in the rooted medium can increase the rooting rate and the fresh weight of roots, and promote the length of root and raise the activities of peroxidase and nitrate reductase significantly. La 3+ has more effect in improving the rooting rate, root length and the activities of peroxidase and less effect in promoting root fresh weight and the activities of nitrate reductase than Eu 3+.

Seed Germination Traits of Loquat (Eriobotrya japonica Lindl.) as Affected by Various Pre-Sowing Treatments (Cutting of Cotyledons, Removal of Perisperm, Moist Chilling and/or Exogenous Application of Gibberellin)

The purpose of this study was to investigate the effects of various presowing treatments on the germinability(final germination percentage)and germination rate of loquat seeds in order to increase seedling production in nurseries(applied research)as well as provide answers for important physiological issues related to loquat seeds and their seed coat(basic research).Three experiments were carried out with various pre-sowing treatments.These treatments included full or partial removal of seed coat(perisperm),partial cutting of cotyledons as well as moist chilling at 5℃ for 13 days and/or soaking the seeds in water or 250 ppm gibberellic acid(GA_(3))solution for 24 h.According to the results,cotyledons excision resulted in delayed germination,regardless of the presence or absence of the seed coat in comparison with the decoated seeds that demonstrated the highest germination rate amongst them.In addition,even the partial excision of seed coats affected positively both the germinability and the germination rate,compared to the control-intact seeds.Furthermore,control-intact seeds had a higher germination percentage when exposed to moist chilling independently of the application or not of gibberellin;while the combination of gibberellin application and moist chilling improved both the percentage and the rate of germination of decoated seeds.In conclusion,the role of perisperm(seed coat)in the germination procedure of loquat seeds seems to be important,indicating the existence of seed coat-imposed dormancy on loquat seeds.Finally,the existence of a mild endogenous embryo-dormancy on loquat is also discussed.

Polyphenol Oxidase Inactivation by Microwave Oven and Its Effect on Phenolic Profile of Loquat (<i>Eriobotrya japonica</i>) Fruit

The objective of this research was investigated the effect of polyphenol oxidase microwave treatment on phenolic composition, antioxidant activity and microstructure of loquat fruit. Phenolic profile of methanolic extracts prepared from fresh, and microwave-treated samples were analyzed. Antioxidant activity was also evaluated by 2,2’-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS?+) and 1,1-diphenyl-2-picrylhydrazyl (DPPH+) methods. In addition, polyphenol oxidase inactivation was carried out using a response surface methodology to establish the optimal conditions of treatment. The phenolic content of fresh mesocarp was 311 ± 0.60 mg gallic acid equivalents (GAE)/100g dry weight (DW) and that of microwave-treated mesocarp was 1230 ± 0.36 mg GAE/100g DW. Total phenolic content of water/ methanol extract significantly increases after microwave treatment rather than methanolic extract of fresh loquat. Five glycoside phenolics were identified by HPLC-DAD-MS as 3-caffeoylquinic acid, 3-p-coumaroylquinic acid, 5-caffeoylquinic acid and quercetin-3-O-sambubioside. Methanolic extract of microwave-treated mesocarp showed higher antioxidant activity than that of fresh mesocarp. Thus, polyphenol oxidase inactivation by microwave energy preserved the integrity of phenolic compounds as well as antioxidant activity in mesocarp extracts prepared from loquat fruit. It was also noted that phenolics were more abundant in the microwaved samples than in the fresh samples.

Selection and Validation of Reference Genes for Normalization of RT-qPCR Analysis in Developing or Abiotic-Stressed Tissues of Loquat (Eriobotrya japonica)

Loquat(Eriobotrya japonica Lindl.)is a subtropical evergreen fruit tree that produces fruits with abundant nutrients and medicinal components.Confirming suitable reference genes for a set of loquat samples before qRT-PCR experiments is essential for the accurate quantification of gene expression.In this study,eight candidate reference genes were selected from our previously published RNA-seq data,and primers for each candidate reference gene were designed and evaluated.The Cq values of the candidate reference genes were calculated by RT-qPCR in 31 different loquat samples,including 12 subgroups of developing or abiotic-stressed tissues.Different combinations of stable reference genes were screened according to a comprehensive rank,which was synthesized from the results of four algorithms,including the geNorm,NormFinder,BestKeeper andΔCt methods.The screened reference genes were verified by normalizing EjLGA1 in each subgroup.The obtained suitable combinations of reference genes for accurate normalization were GAPDH,EF1αand ACT for floral development;GAPDH,UBCE and ACT for fruit setting;EF1α,GAPDH and eIF2B for fruit ripening;ACT,EF1αand UBCE for leaves under heat stress;eIF2B,UBCE and EF1αfor leaves under freezing stress;EF1α,TUA and UBCE for leaves under salt stress;ACT,EF1αand eIF2B for immature pulp under freezing stress;ACT,UBCE and eIF2B for immature seeds under freezing stress;EF1α,eIF2B and UBCE for both immature pulp and seeds under freezing stress;UBCE,TUB and TUA for red-fleshed fruits under cold-storage stress;eIF2B,RPS3 and TUB for white-fleshed fruits under coldstorage stress;and eIF2B,UBCE and RPS3 for both red-and white-fleshed fruits under cold-storage stress.This study obtained different combinations of stable reference genes for accurate normalization in twelve subgroups of developing or abiotic-stressed tissues in loquat.To our knowledge,this is the first report to obtain stable reference genes for normalizing gene expression of abiotic-stressed tissues in E.japonica.The use of the three most stable reference genes could increase the reliability of future quantification experiments.

腐植酸水溶肥对枇杷幼苗生长的影响

为研究叶面喷施腐植酸水溶肥对枇杷[Eriobotrya japonica(Thunb.)Lindl.]幼苗生长的影响,以大五星枇杷幼苗为材料,叶面喷施不同稀释浓度(700、1 000、1 300、1 600倍)的腐植酸水溶肥,以喷施等量清水为对照(CK),测定腐植酸水溶肥对枇杷幼苗生物量、光合色素含量和抗氧化酶活性的影响。结果表明,叶面喷施不同浓度的腐植酸水溶肥提高了枇杷幼苗茎杆和叶片生物量、光合色素含量,且均以稀释1 000倍效果最好。稀释1 000、1 300倍腐植酸水溶肥处理提高了枇杷幼苗过氧化氢酶(CAT)、过氧化物酶(POD)和超氧化物歧化酶(SOD)活性。枇杷幼苗茎杆、叶片生物量与叶绿素a、叶绿素b、类胡萝卜素含量、CAT活性、POD活性和SOD活性呈正相关。因此,叶面喷施腐植酸水溶肥能够促进枇杷幼苗生长,且以稀释1 000倍最佳。

千岛湖地区不同蜜蜂传粉对枇杷增产提质效果的评价

应用中华蜜蜂和意大利蜜蜂为浙江省千岛湖地区的枇杷传粉,评估2种蜜蜂传粉对枇杷果实品质和产量的影响。结果表明,枇杷花期不同时间2种蜜蜂访花密度差异较大,其中9:30~9:50时,中华蜜蜂的访花密度(10.35±2.15只/100朵花)显著大于意大利蜜蜂的访花密度(5.00±1.39只/100朵花)(P<0.05),其余时段2种蜂的访花密度差异不显著。经2种蜜蜂传粉的枇杷果实维生素C含量、总糖、糖酸比、单果重及果形指数等指标均显著优于隔离对照区的枇杷果实(P<0.05),在改善枇杷果实品质方面2种蜜蜂传粉效果基本一致。大田亩产量调查结果显示,中华蜜蜂传粉区的平均枇杷亩产量略高于意大利蜜蜂传粉区,但二者之间差异不显著(P>0.05)。本研究认为2种蜜蜂传粉均达到枇杷增产提质的效果,其中中华蜜蜂更适合为山区冬季开花的枇杷传粉。

低温胁迫下油菜素内酯对枇杷幼果生理特性的影响

对枇杷(Eriobotrya japonica LindL.)幼果使用0、0.5、1.0、1.5、2.0 mg/L外源油菜素内酯预处理后,于-3℃条件下低温处理24 h,研究低温胁迫下油菜素内酯对枇杷幼果相对电导率、抗氧化酶活性和渗透调节物质含量等生理指标的影响。结果表明,低温处理使枇杷幼果的相对电导率显著升高,冻害指数升高至90.5,但随油菜素内酯浓度的增加呈下降趋势;与0 mg/L相比,枇杷幼果的SOD、POD、CAT、APX酶活性和可溶性糖、可溶性蛋白质、脯氨酸含量在油菜素内酯的作用下得到不同程度的提高,并均与其相对电导率呈极显著负相关。这表明在低温胁迫下油菜素内酯可以通过调控抗氧化酶活性,提高渗透调节物质含量,增强枇杷幼果的耐寒性,并在油菜素内酯浓度为2.0 mg/L时枇杷幼果的耐寒性最佳。

大棚枇杷上木虱发生规律及5种杀虫剂的防治效果

浙江兰溪枇杷是农业农村部农产品地理标志产品,随着大棚设施普及,以木虱为主的害虫发生日益严重。在浙江省兰溪市大棚枇杷主产区调查大棚和露天枇杷上木虱的发生规律,比较不同地点木虱发生情况,并对大棚枇杷上木虱开展阿维菌素·双丙环虫酯干悬浮剂等5种药剂防控试验。结果表明,浙江省兰溪市女埠街道大棚枇杷上木虱若虫一年高发期3个,露天枇杷上木虱若虫一年高发期2个,其中大棚枇杷上木虱高发期相比露天早半个月左右。大棚和露天栽培的枇杷上木虱虫口发生量差异极显著(p<0.01),大棚枇杷上木虱平均虫量4.8头/枝,是露天的5.3倍;4个调查地点中女埠街道枇杷上木虱发生最重。75 g/L阿维菌素·双丙环虫酯干悬浮剂15 mL/667 m^(2)处理药后5、10、15 d的防效较高,分别为91.03%、90.88%和86.28%,显著高于5%阿维菌素微乳剂4000倍液,其药后15 d防效仅为46.34%。阿维菌素·双丙环虫酯防效好,使用安全,可在实际生产中推广应用。

枇杷PYL基因家族鉴定及表达分析

脱落酸(ABA)是调节植物生长发育与非生物胁迫的重要激素之一,ABA受体Pyrabactin Resistance 1(PYR)-like(PYL)于信号通路的顶端发挥核心调控作用,但枇杷Eriobotrya japonica的PYL基因家族尚未鉴定。本研究通过生物信息学分析鉴定出枇杷12个PYL基因家族成员,枇杷PYL蛋白分子量为10.47~29.12 kDa,且主要定位于细胞质。枇杷PYL基因家族成员分布在8条染色体上,进化树分析显示PYL基因家族包含三个亚家族,且枇杷EjPYL1与拟南芥AtPYL13亲缘关系较近。枇杷PYL蛋白含有1~6个motif,其中motif3为家族成员共有。枇杷PYL基因家族成员在物种内存在共线性,与拟南芥的物种间共线性有2对。顺式作用元件分析发现枇杷PYL基因启动子含有低温、干旱、光以及激素响应元件。枇杷幼果转录组分析表明,果肉和种子中EjPYL6和EjPYL8基因受低温诱导上调表达。该研究结果将为枇杷ABA信号转导途径的解析和抗冻新品种培育提供理论依据。

枇杷叶抗炎和止咳作用研究

目的 研究枇杷叶抗炎和止咳作用的有效成分 ,为制定枇杷叶的质量标准提供科学可靠的实验依据。方法采用二甲苯致小鼠耳肿胀法、二氧化硫引起小鼠咳嗽和枸橼酸喷雾致豚鼠咳嗽模型 ,观察枇杷叶不同分离部分及单体成分的抗炎及止咳作用。结果 P2和 P3部分对小鼠耳肿胀法所致的炎症具有较好的对抗作用 ,且 P2部分具有明显的止咳作用。P2部分中的枇杷苷 ,P3部分中的总三萜酸和熊果酸均具有良好的抗炎、止咳效果。结论 枇杷苷 、熊果酸和总三萜酸是枇杷叶抗炎、止咳作用的主要成分 。

枇杷叶中三萜类成分的研究

采用色谱方法对枇杷叶成分进行分离纯化,并依据理化性质和波谱数据鉴定化合物的结构。结果从枇杷叶乙醇提取物中分离到13个化合物,经鉴定为三萜和脂肪醇类物质,其中三萜:乌苏酸(1)、蔷薇酸(2)、齐墩果酸(3)、2α,19α-二羟基-3-羰基-12-烯-28-乌苏酸(4)、2α,3β,13β-三羟基-11-烯-28-乌苏酸(5)、2α-羟基白桦脂酸甲酯(6)、科罗索酸甲酯(7)、乌苏醇(8)、科罗索酸(9)、2α,3α,19α-三羟基-12-烯-28-齐墩果酸(10)、β-谷甾醇(11)、2α,3α,19α,23-四羟基齐墩果酸(12);脂肪醇:正二十一烷醇(13)。化合物5、6、8、12和13是首次从枇杷属植物中分离得到。

‘早钟6号’枇杷主要营养元素含量的年周期变化

‘早钟 6号’枇杷叶片中较高的N、P营养水平有利于花芽分化 ;叶片中营养元素含量次序为Ca >N >K >Mg >P ,花穗为K >N >Ca >Mg >P ,果实为K >N >Ca >P >Mg ;认为早钟 6号枇杷属喜Ca、喜K果树 ,生产上应重视Ca肥和K肥的施用 ;枇杷叶片营养分析采样以第一次夏梢叶片为宜 ,初步确定叶片营养诊断的采样适期为 9月。

NaHSO_3对枇杷和毛叶枣叶片光合速率的促进作用

研究了不同浓度的NaHSO3及其与KCl和6-BA配合施用后对田间枇杷和毛叶枣离体叶片光合作用的影响。结果表明:低浓度(2～8mmol/L)的NaHSO3对光合速率有促进作用,作用效果长达6d;高浓度(16mmol/L)的NaHSO3对光合速率促进作用不明显。NaHSO3配合1mmol/LKCl和5mmol/L6-BA施用,对光合速率的促进效果更明显。

枇杷叶中三萜类化学成分的研究

目的:研究枇杷叶的化学成分。方法:采用硅胶、ODS柱、制备薄层及制备液相法分离枇杷叶中的化学成分,运用化学和有机波谱分析方法确定化合物结构。结果:从枇杷叶中分离得到9个三萜类化合物,经鉴定分别为白桦脂酸甲酯(Ⅰ)、齐墩果酸(Ⅱ)、乌苏酸(Ⅲ)、2α-羟基齐墩果酸甲酯(Ⅳ)、科罗索酸甲酯(Ⅴ)、2α-羟基齐墩果酸(Ⅵ)、科罗索酸(Ⅶ)、委陵菜酸(Ⅷ)、蔷薇酸(Ⅸ)。结论:化合物Ⅰ、Ⅴ为首次从该植物及该属植物中分离得到。

枇杷花三萜皂甙成分的研究

目的 研究枇杷花的化学成分。方法 通过化学方法和波谱分析 (IR,MS,1 HNMR,1 3CNMR)。从枇杷〔Eriobotrya japonica(Thunb) L indl〕的花中经乙醇提取分离三萜皂甙元类成分。结果 共获得 4个三萜皂甙元类成分 ,并确定它们的化学结构分别为 : 齐墩果酸 (oleanolic acid) , 熊果酸 (ursolic acid) , 2 α,3α,19α-三羟基熊果 - 5 ,12 -二烯 - 2 8-酸 (2 α,3α,19α- trihydroxyurs- 5 ,12 - dien- 2 8- acid) , 2 β,3β,2 3α-三羟基齐墩果 - 12 -烯 - 2 8-酸(2 β,3β,2 3α- trihydroxyolean- 12 - en- 2 8- acid)。结论 化合物 ～

GA_3诱导的单性结实‘宁海白’白沙枇杷糖代谢的研究

研究了花蕾期GA30·5、1·0g·L-1处理诱导的单性结实‘宁海白’无核白沙枇杷的糖积累与代谢特性。结果表明:两种浓度的GA3诱导的无核枇杷中,其蔗糖、葡萄糖、果糖和总糖的含量均低于正常有核果,并随处理浓度的增加而下降,其中成熟时处理与对照之间各种糖的含量差异达到显著水平。不同浓度GA3处理后,果实中蔗糖代谢酶(SS、SPS、转化酶)、己糖代谢酶(HXK、FRK)和山梨醇脱氢酶(SDH)等的活力均低于正常有核果,并随处理浓度的增加而下降。上述结果表明,GA3诱导无核,导致果实糖代谢酶活力的下降,是无核枇杷果实含糖量下降的主要因子。

低温等因素对枇杷种质离体保存的影响

初步研究了低温条件下枇杷(EriobotryajaponicaLindl.)试管苗离体保存的效果。结果表明:在常温(25℃)条件下,枇杷试管苗生长速度快,保存时间短,10个月后全部死亡;而在低温条件下,试管苗生长缓慢,保存时间较长。在11℃和15℃条件下,品种'解放钟'和'早钟6号'1a后的死亡率分别为20%、25%和15%、20%,表明低温有利于枇杷种质资源的离体保存。在试管包扎物的选择中,白纸+塑料薄膜和锡箔纸等材料封口对保存有利,而棉花、软木塞、橡皮塞等封口材料不利于试管苗的保存。

枇杷花系统溶剂提取物抑菌作用研究

研究枇杷花系统溶剂提取物抑菌的作用。实验以苯甲酸钠为对照组,分别研究了枇杷花石油醚、氯仿、乙酸乙酯、正丁醇、丙酮、95%乙醇和水的提取物对大肠杆菌、金黄色葡萄球菌、白色念珠菌的抑菌效果,并对不同提取物进行薄层色谱分析。结果表明,枇杷花的提取液对细菌、真菌均有抑制作用,并优于对照组。正丁醇、丙酮提取物对金黄色葡萄球菌抑制能力优于其他溶剂;石油醚提取物对大肠杆菌抑制能力优于其他溶剂;石油醚、乙酸乙酯提取物对白色念珠菌抑制能力优于其他溶剂。

NaNO2-Al（NO3）3-NaOH比色法测定枇杷花水提液中总黄酮含量

目的：建立紫外-可见分光光度法测定枇杷花水提液中总黄酮的含量,为枇杷花新食品原料的申报奠定基础。方法：以芦丁为对照品,采用NaNO2-Al（NO3）3-NaOH显色反应体系,对显色剂的浓度与用量进行考察,筛选出最佳显色条件,在505 nm处用紫外-可见分光光度法测定样品中总黄酮的含量。结果：采用该方法可以准确地测定枇杷花水提液中总黄酮的含量,总黄酮在16.4898.88μg/mL范围内线性关系良好,回归方程为y=6.470 2x-0.003 7,R2=0.999 4,样品平均回收率为95.56%,RSD为3.1%。结论：该方法简便、快捷、准确,可作为枇杷花水提液总黄酮含量测定的方法。

胶束毛细管电泳测定枇杷叶中熊果酸和齐墩果酸的含量

目的:建立测定枇杷叶中熊果酸和齐墩果酸含量的毛细管电泳方法。方法:采用胶束毛细管电泳法(MECC),未涂层弹性石英毛细管(75 μm×60 cm,有效长度50 cm);压力进样,压力:3.0×10~3 Pa;进样时间:5 s;分离电压:24 kV;柱温:25℃;检测波长:200 nm;运行缓冲液:30 mmol·L^(-1) SDS,20 mmol·L^(-1)羟丙基-β-环糊精,5%乙腈,40 mmol·L^(-1)硼砂溶液(pH9.4);运行时间:30 min。结果:测得熊果酸和齐墩果酸的线性范围分别为42.5～680 μg·mL^(-1)(r=0.9989)和33.0～660μg·mL^(-1)(r=0.9994),平均回收率(n=5)分别为98.1%(RSD 为3.3%)和97.8%(RSD 为2.9%)。结论:方法准确可靠,适合于枇杷叶中熊果酸和齐墩果酸的含量测定。