Astaxanthin, a ketocarotenoid with superior antioxidant properties compared with other carotenoids, is increasingly demanded in feed, food, and pharmaceutical industries. This study aimed to engineer apple (Malus domestica Borkh.) callus and tobacco (Nicotiana benthamiana Domin) leaf for sustainable production of natural (3S,3'S)-astaxanthin. A plant expression vector, pRI101-Flag-BHY-BKT-PSY1-CrtI (BBPC), was constructed to deliver astaxanthin biosynthetic genes PSY1, CrtI, BHY and BKT genes into 'Orin' apple callus and N. benthamiana leaf. Transgenic apple calli synthesized 40.4 mu g/g dry weight (DW) astaxanthin, representing 53.5 % of total carotenoids. Transiently transformed N. benthamiana leaves produced 732.8 mu g/g DW astaxanthin,accounting for about 18.2 % of total carotenoids. Furthermore, supplementing tomato juice (20 mL/L) and carrot juice (30 mg/L) increased astaxanthin levels in calli by 1.7-fold. Moreover, astaxanthin-rich tissues exhibited 2-4 fold higher antioxidant activity (P < 0.05) than controls. Acute toxicity assays in mice revealed no adverse effects. These findings demonstrate the potential of engineered apple callus and N. benthamiana leaf as scalable sources of natural (3S,3'S)-astaxanthin for industrial applications.

Flavonoids are one of the main pigments in horticultural fruit. The AP2/ERF transcription factors play various roles in higher plants, and thus, MdAP2–34 has been reported to regulate carotenoid accumulation in apple. However, it is unclear whether light interferes with the flavonoid accumulation in apple fruit flesh. Here, we found that apple of ‘Benin Shogun’ fruit flesh had higher levels of flavonoids under light treatment than under dark treatment, with an increase of 48% at 15 days after harvest (DAH) and 39% at 30 DAH. The MdAP2–34 transcript levels exhibited a highly positive correlation with the flavonoid content of ‘Benin Shogun’ fruit under dark and light treatments. Furthermore, the structural genes of flavonol biosynthesis, including MdCHS, MdCHI, MdF3′H, and MdFLS, were positively associated with flavonoid contents of fruit and were upregulated in MdAP2–34-OVX apple calli. The MdAP2–34 stimulated flavonoid accumulation via the flavonoid biosynthetic pathway by directly binding to and activating the MdF3′H promoter activity. Our findings demonstrate the use of multiple MdAP2–34-mediated functions in understanding the complex mechanisms of flavonoid synthesis in apple, which may also be useful in apple breeding.

ABSTRACT This study investigated the impact of compost amendment on fungal structure of the replanted apple orchard in comparison with the original orchard without compost improvement. The V4 region of the fungal 18S rRNA gene was sequenced using high-throughput 454 pyrosequencing, and the data were analyzed using the Mothur pipeline. The results showed that the compost amendment modified the fungal community structure at all soil depths (0–20 and 20–40 cm) and sites (in-row and inter-row) considered, such as the relative abundances of Vampyrellidae (16.2%), Trichocomaceae (14.2%) and Rhizaspididae (2.5%) were increased while that of Glissomonadida (15.4%), Eugregarinorida (4.0%) and Thecofilosea (3.4%) were decreased. Overall, our findings show that compost amendment altered the fungal community compositions in replanted orchard soil.

黄河三角洲位于山东省东北部,是世界上暖温带最年轻的湿地生态系统.由于成陆前受海水长期浸渍,地下水位较高,且有较高的矿化度,土壤年蒸发量大,因而形成了大面积的盐碱地.土壤盐渍化严重阻碍当地传统农业经济的发展,为加强盐碱地综合利用和探索适于盐碱地的果树栽培模式,本文结合国家行业标准及盐碱地葡萄栽培技术特点,从园地选择、大棚设计、苗木栽植、土壤改良、花果管理、温湿度调控及病虫害防治等设施葡萄栽培一系列关键技术出发,提出了黄河三角洲大棚葡萄标准化栽培技术规程,旨在为当地葡萄产业升级和可持续健康发展提供理论参考.

花青素是植物中最广泛存在的次生代谢物之一,是不同颜色花和果实色泽的关键色素.花青苷通常在细胞质中合成,于液泡中积累.虽然花青苷的生物合成途径己被深入研究,但它们从细胞质到液泡的转运以及显示红色的机制仍然不清楚.本试验通过RACE技术成功克隆出富士苹果及其芽变中MdVAMPcDNA序列的总长度.该基因cDNA序列全长994bp,具有660 bp的开放阅读框(ORF),编码219个氨基酸.同源性分析表明,MdVAMP基因与沙梨同源性最高.定量PCR结果表明,MdVAMP基因在‘长富2’苹果及芽变果实摘袋后表达趋势与其花青苷含量变化相一致,芽变果实中MdVAMP基因表达量和花青苷含量均显著高于‘长富2’苹果.进一步在不同富士品种中研究发现,MdVAMP基因表达水平与花青苷含量密切相关.亚细胞定位结果显示MdVAMP主要定位于细胞质.该研究表明,MdVAMP基因可能参与了花青苷的转运和积累,这为进一步研究苹果果皮着色和花青苷转运提供了一定的科学基础.

以‘沾冬2号’冬枣为试材,在冬枣果实套无纺布果袋条件下,测定了枣果生长期间袋内外温度及空气相对湿度,并分析了白熟期、脆熟期和完熟期冬枣外观品质及相关指标,研究了无纺布果袋对‘沾冬2号’冬枣微域环境和果实品质的影响,以期为应用无纺布果袋套袋技术生产优质冬枣提供技术支撑.结果表明:套袋导致袋内温度升高、湿度下降;袋内最高温度比袋外最高温度高1～2℃.套袋后冬枣果实枣皮光滑、机械损伤较小,并且在白熟期、脆熟期、完熟期与不套袋相比果皮亮度分别提高了3.06％、16.46％、9.88％;果实可溶性糖含量分别降低了0.21％、1.99％、0.94％;可滴定酸含量分别升高了42.85％、13.79％、9.20％;维生素C含量分别降低了0.30、0.17、0.09 mg·g-1,但差异均未达显著水平.套袋能明显改变冬枣果实微域环境,改善枣果外观品质,对其内在品质无明显影响.

Apples may suffer sunburn during growing season because of photooxidation damage. Astaxanthin have a stronger antioxidant activity against photooxidation compared to its precursors zeaxanthin and beta-carotene. Here, we transferred beta-carotene ketolase (bkt) and beta-carotene hydroxylase (crtR-B), two genes encoding key enzymes for astaxanthin biosynthesis, into "Brookfield Gala" apple plants using pCAMBIA1302-bkt-crtR-B via transformation mediated by Agrobacterium tumefaciens EHA105. We aimed to confer a stronger antioxidant ability against sunburn to apple plants by regulating metabolic pathways of carotenoids. RT-PCR and qPCR analysis confirmed that bkt and crtR-B genes were expressed simultaneously in transgenic plants. Southern blotting analysis confirmed that the transgene was stably integrated and determined transgene copy number. Transgenic plants synthesized novel ketocarotenoids, resulting in an accumulation of astaxanthin up to 12.06 mu g/g and of canthaxanthin up to 6.38 mu g/g per leaf fresh weight. The light antioxidant ability of transgenic plantlets was stronger than that of non-transgenic ones in high light conditions. We speculate that the transgenic apples accumulating astaxanthin are resistant to photooxidation and can therefore prevent sunburn.

葡萄砧木品种类型多样,抗性各异,在生产中起着重要的作用,对接穗品种的生长发育和浆果品质产生重要影响;葡萄砧木的嫁接效应也与砧木类型、接穗品种、立地条件、土壤和肥水管理措施有关.本文根据近年来国内外葡萄砧木的研究成果,介绍了葡萄砧木的主要种类品种,并对葡萄砧木的抗性研究方法和砧木选择性应用做了简要的综述;进一步概述了葡萄砧木对接穗生长发育、产量和果实品质的影响;各地需根据土壤和气候等特点因地制宜地进行葡萄砧木品种区域化研究与应用,以促进我国葡萄产业的健康和可持续性发展.

Compost amendment reportedly improved apple tree growth in replant soils. However, its effects should be evaluated at different soil depths and locations. This study investigated the impact of soil improvement with compost on soil physicochemical properties and bacterial community structure of a replanted apple orchard in comparison with the original orchard without compost improvement. The V1-V3 region of the bacterial 16S rRNA gene was subjected to high-throughput 454 pyrosequencing, and data were analyzed using the Mothur pipeline. The results showed that the soil improvement benefited tree growth and fruit quality during the study period. The compost amendment markedly increased tree height and stem diameter by a range of 6.1%-21.0% and 4.0%-14.0%, respectively. Fruit yield (9.5%), average weight (9.6%), and soluble solid content (5.6%) were also increased by compost amendment compared to those of the unimproved treatment. The pH, organic matter, and available N, P, and K contents were significantly increased by 5.7%-21.9%, 0.2%-62.9%, 9.3%-29.3%, 36.7%-64.5%, and 17.2%-100.3% in the compost improved soil. The pyrosequencing data showed that the soil improvement changed the bacterial community structure at all soil depths (0-20 cm and 20-40 cm) and locations (in-row and inter-row) considered; e.g., the relative abundance of Proteobacteria (20.2%), Bacteroidetes (2.5%), and Cyanobacteria (1.0%) was increased while that of Chloroflexi (5.5%), Acidobacteria (5.2%), Nitrospirae (4.5%), Gemmatimonadetes (3.8%), and Actinobacteria (1.8%) was decreased. The relative abundance of some dominant genera Burkholderia (2.3%), Pseudomonas (1.0%), and Paenibacillus (0.5%) were enhanced in the compost improved soil. Moreover, other dominant genera such as Nitrospira (6.4%), Gemmatimonas (2.2%), and Phenylobacterium (0.3%) were reduced by the application of compost. Our results indicate that soil improvement benefits the growth of tree and fruit quality, and is likely mediated by increased soil pH, organic matter, and available nutrient contents and beneficial bacterial community composition.

Anthocyanin which has been found many kinds of health care effects is one of the most important pigments for the fruit,In order to explore the relationship between anthocyanin's degradation and pectate lyase in ripening of peach,we used the reportorial pectate lyase gene sequence of peach to design special primers and clone the pectate lyase gene (PpPL) who has an open reading frame of 1 242 bp and encode 413 amino acids,We studied the expression of PpPL in the different maturity and tissues of the peach by qRT-PCR,The results showed that the PpPL gene in 'Wutao' showed an increasing trend with the fruit growing and anthocyanin degradation and a significant increase when fruit started soft,There was a difference in gene PpPL expression among different tissues,The relative expression at the same maturity in fruit pulp＜leaf＜fruit peel＜ stems,so we theorize that PpPL may be involved in the degradation of anthocyanins.

随着网络技术的发展,教学手段和教学方法日益丰富.在传统的教学基础上,网络技术为教学过程搭建了新的平台,网络教学平台的建设和有效利用直接影响教学效果.应用网络平台进行遗传学教学,不仅提高教学效率,而且也便于学生自主和延展性学习.经多年实践与探索表明,网络教学是遗传学教学有效的辅助手段之一,具有传统教学无法比拟的优势,它实现了"教与学"信息的电子化和网络化,强化了教学资源的共享,促进了师生互动.该文就遗传学网络教学平台的设计原则、内容框架和应用维护等进行了探讨,为遗传学理论教学的创新模式探索提供了新的思路.

This research used‘Yanfu 8’that was red bud mutation variety from‘Fuji’as the experimental mate-rial, cloned UV-B receptor gene named MdUV R 8 by RT-PCR from fruit peel. The results showed that the full-len-gth of UV R8 gene open reading frame (ORF) was 1 359 bp in size and encoded 452 amino acids residues (Mw=48.487 kD, pI=5.56). Conservative protein domain analysis showed that apple UVR8 protein contains seven RCC1 domain structure, moreover, protein secondary structure prediction revealed that it contains 10 alpha helixes, 16 be-ta folds and 41 beta turns. Amino acids homology comparison analysis indicated that sequence had 69.54%~88.94%similarity with those of other reported plants. Nucleotide cluster analysis showed that UV R8 from Pyrus bretsch-neideri was clustered together with apple's UV R8 firstly, and followed by which from Prunus mume. This research laid a foundation for further research of the molecular mechanism of photoreceptor response to light in apple.

以葡萄品种“巨峰”为试材，研究黄河三角洲不同果园土壤盐分、pH值和养分等状况及其对葡萄生长发育的影响。结果表明，该地土壤含盐量与电导率有显著相关性。平地和台田果园不同深度土层的土壤含盐量和养分存在差异，表层土壤（0～20 cm）对盐分具有积聚作用，并呈现季节性变化。与平地果园相比，台田果园各层土壤容重相对较低，脱水后其最大值出现时间较迟，且增加幅度不同；土壤紧实度显著低于平地果园，但在水平方向连续性不强；pH值和氮磷钾等养分状况优于平地果园。在对葡萄树体生长发育和果实品质的影响方面，平地果园葡萄明显表现生长势偏弱，果实发育不良。

A full-length glutathioneS-transferase gene namedMdGSTF3 was cloned with RT-PCR and RACE-PCR fromMalus domestica cv. ‘Fuji’ peel. The GenBank accession number was KP234026.MdGSTF3was 868 bp in length including 49 bp of 5' untranslated region, 151 bp of 3' untranslated region and 26 bp polyA. It had a coding sequence (CDS) of 642 bp, encoding 213 amino acid residues. This predicted polypeptide includ-ed 19 kinds of amino acids except for cysteine, with the molecular mass of 23.993 kDa, isoelectric point of 6.17. Real-time lfuorescent quantitative PCR analysis showed that the expression level ofMdGSTF3 in stems, leaves, lfowers of striped-red ‘Fuji’ was higher than that of the mutant blushed-red ‘Fuji’, but the expression level of MdGSTF3 in peel and pulp of blushed-red ‘Fuji’ was higher than that of striped-red ‘Fuji’. The most relative expression quantity ofMdGSTF3 from peel and pulp of blushed-red ‘Fuji’ were 46.6% and 22.1% higher than that of striped-red ‘Fuji’ respectively. The results indicated that the expression level ofMdGSTF3was closely related with anthocyanin accumulation and distribution of apple peel.

Zhonghuashoutao peach fruits were treated with salicylic acid(SA)at the concentration of 1. 25,2. 50, 5. 00 mmol/L and effect of SA on browning in peach fruit during storage under (1 ± 0. 5) ℃ and RH(90 ± 5)% was studied. The experiment items respectively were peach flesh color and luster, firmness, polyphenoloxidase and li-poxygenase activity,the content of hydrogen peroxide and malondialchehyche,production rate of superoxide anion in peach pulp tissue related with fruit browning. The results showed that SA not only could retard the rate of peach fruit firmness descending to a certain degree,but also reduce the activity of polyphenoloxidase and lipoxygenase. SA also could reduce production rate of superoxide anion and enhance content of hydrogen peroxide at the earlier stage of storage. It could remain higher balance of reactive oxygen species metabolism. In addition,SA acid might alleviate the harm of malondialchehyche on cell membrane and keep its stability. SA reduced browning index obviously com-pared to control and restrained the development of fruit browning,but all SA treatments had no significant difference on browning index .

The change of gene expression level during coloring period is crucial for the formation of apple fruit quality. The selection of a suitable reference gene is an important factor for accurate gene expression analysis by real-time lfuorescence quantitative PCR (qRT-PCR). In this study, peels of apple (Malus domestica cv.‘Fuji’) were taken as materials sampled at different time during coloring stage. The expression level of four commonly used housekeeping genesβ-actin, EF-1α, GAPDH and 18S rRNA were studied by qRT-PCR, and reliable refer-ence gene were screened to use for gene expression during coloring stage of fruit by geNorm and NormFinder software. The results showed that the expression level of EF-1αwas highest and most stable, followed by 18S rRNA. However, the expression level ofβ-actin and GAPDH were relatively low. Meanwhile, the expression of lfavonoid-4-reductase gene (MdDFR) in the pathway of apple anthocyanin biosynthesis was analyzed, and the results indicated that the variation tendency of MdDFR was exactly consistent and presented normal distribu-tion using EF-1αand 18S rRNA as reference gene. Therefore, EF-1αand 18S rRNA are suitable reference genes and could be used to normalize mRNA levels in peel tissues of apple during coloring period.

We used ‘whangkeumbae’ pear fruits as experimental materials and studied the effect of different package methods on some postharvest physiological index of ‘whangkeumbae’ pear under CA storage. The package methods mainly included PE freshness-maintaining bags which were 0.013-mm and 0.02-mm-thick, double-layer paper bag and the unpacked respectively. Fruit firmness, total soluble solids, rate of ethylene production, rate of respiration, vitamin C (Vc) content, chlorophyll content, index of browning and decay, rate of good fruit and weight loss were studied during storage. The results showed that 0.013-mm thick freshness-maintaining bag could produce better storage results on above-mentioned indexes than the others. It mainly showed that it could prevent fruit firmness from declining and delay fruit softening, maintain higher total soluble solids, reduce and postpone ethylene production and respiration peak, and fresh Vc and chlorophyll in peel lost little. To some extent, it could restrain fruits browning and lessen the rotten ratio of fruits. Rate of good fruit and weight loss were 93.63% and 4.32% after stored for 180 days.

Taking Prunus persica 'Zhonghuashoutao' fruits as material,peach fruits were harvested with four different maturity degrees and stored for 7 weeks.It was performed to investigate the postharvest changes in firmness,soluble solids content,titratable acid,respiration rate,ethylene production,cell membrance permeability,degree of fruit browning and sensory quality so as to find the best harvesting dates suiting to be stored at low temperature(1±0.5)℃,RH(90±5)%.The results showed that the effects of storage harvesting on October 25 obviously was superior to the others at firmness,soluble solids content etc.At the same time applying sensory evaluation in fruit quality later storage period proved that the effects of different harvesting dates on 'Zhonghuashoutao' peach fruit during storage were October 25th,October 20th,October 30thand October 15th according to its priority.

虾青素是一种氧化型酮式红色类胡萝卜素,具有更强的抗光氧化能力.将β-胡萝卜素酮化酶(虾青素生物合成的关键酶)基因bkt构建入表达载体pCAMBIA1301中,获得植物表达载体p1301-bkt,转化根癌农杆菌EHA105,获得工程菌,以黄肉苹果‘Brookfield Gala’无菌试管苗叶片为受体,进行遗传转化.筛选压确定结果表明:‘Brookfield Gala’对潮霉素(Hyg)很敏感,叶片再生最佳Hyg选择压为3 mg·L-1,试管苗增殖为4 mg·L-1,生根为2 mg·L-1;头孢霉素(Cef)浓度≤400 mg·L-1时对叶片再生芽数的影响不明显.GUS染色、PCR和RT-PCR检测结果表明,有8株转基因植株整合bkt基因并获得了表达,其表型有红色产生;转基因植株类胡萝卜素的HPLC测定显示,虾青素和角黄素在叶片中的积累量达到2.85和1.79 μg·g-1.本研究结果显示有望通过调控代谢途径,在苹果中合成虾青素,提高果实自身的抗光氧化能力,防止日灼.

'Fruit tree cultivation' is an important professional content-based course of horticulture specialty in agricultural universities.In the previous teaching activities,students could hardly apply their learned theory to practice because of the less access to practical training.To meet the demand of training aim,fruit production practice should be added to 'fruit tree cultivation' curriculum,along with the adoption of student-oriented teaching model.Besides,to cultivate talents with broad knowledge and professional skills,the present paper suggests optimizing the teaching content and method,reforming the teaching evaluation system,enhancing teachers' teaching standards,and setting up the bases for practical teaching.