Root demography in kiwifruit (Actinidia deliciosa)

A rhizotron was used to study fine-root demography in mature vines of kiwifruit (Actinidia deliciosa). The vines were grown in a deep, well drained, silt loam and received normal orchard management. Roots were measured from 10 to 160cm depth at biweekly intervals for 2 years. After an initial phase of rapid colonisation of the repacked soil behind the rhizotron windows, the total length of visible roots per vine remained quite steady. This apparent stability of the total belied fast and sustained localized turnover of the fine roots at all soil depths. Fifty-one per cent of the roots survived ≤28d, 69% died at an age ≤56d and only 8% survived >252d. For each year, the cumulative length of roots grown was equivalent to about 2·75 times the maximum net length of roots visible. These may be the largest annual rates of root turnover yet reported. This has important ramifications for the carbon balance, mineral nutrition and water relations of the plant.     

Development of a 135K SNP genotyping array for Actinidia arguta and its applications for genetic mapping and QTL analysis in kiwifruit

Kiwifruit (Actinidia spp) is a woody, perennial and deciduous vine. In this genus, there are multiple ploidy levels but the main cultivated cultivars are polyploid. Despite the availability of many genomic resources in kiwifruit, SNP genotyping is still a challenge given these different levels of polyploidy. Recent advances in SNP array technologies have offered a high-throughput genotyping platform for genome-wide DNA polymorphisms. In this study, we developed a high-density SNP genotyping array to facilitate genetic studies and breeding applications in kiwifruit. SNP discovery was performed by genome-wide DNA sequencing of 40 kiwifruit genotypes. The identified SNPs were stringently filtered for sequence quality, predicted conversion performance and distribution over the available Actinidia chinensis genome. A total of 134 729 unique SNPs were put on the array. The array was evaluated by genotyping 400 kiwifruit individuals. We performed a multidimensional scaling analysis to assess the diversity of kiwifruit germplasm, showing that the array was effective to distinguish kiwifruit accessions. Using a tetraploid F1 population, we constructed an integrated linkage map covering 3060.9 cM across 29 linkage groups and performed QTL analysis for the sex locus that has been identified on Linkage Group 3 (LG3) in Actinidia arguta. Finally, our dataset presented evidence of tetrasomic inheritance with partial preferential pairing in A. arguta. In conclusion, we developed and evaluated a 135K SNP genotyping array for kiwifruit. It has the advantage of a comprehensive design that can be an effective tool in genetic studies and breeding applications in this high-value crop.     

The development and effects of nitrogen deficiency in field-grown kiwifruit (Actinidia deliciosa) vines

The development and effects of nitrogen (N) deficiency in kiwifruit (Actinidia deliciosa Hayward) vines planted at three densities (25.0, 12.5 and 8.33 m² vine^–1) were examined in a long term (1982 to 1989) field experiment in which N was applied at rates from 0 to 200 kg N ha^–1 year^–1. The rate of applied N significantly affected leaf N concentrations every year from 1985 onwards, and the average leaf N concentrations declined throughout the experiment. Fruit N concentrations varied significantly with the level of applied N as early as 1986. The average fruit N concentrations varied strongly between years, and were inversely proportional to the fruit number (per m²), indicating that, after fruit set, growth of individual fruit was relatively insensitive to the vine N status. Effects of N supply on fruit yields resulted mostly from changes in fruit number (per m²). For vines planted at the high density, fruit yields responded significantly to the level of applied N each season from 1986 onwards. In any year, maximum fruit yields for vines planted at the high density were associated with leaf N concentrations (20 weeks after bud burst) of at least 1.8 mmol g^–1. For vines planted at low density, significant yield responses to the level of applied N were not recorded until 1988, and maximum yields in that year were associated with leaf N concentrations of at least 1.4 mmol g^–1. The delayed expression of effects of N deficiency on fruit yields for vines planted at low density appeared to follow a shift in partitioning of resources in favour of fruit growth. This shift in partitioning did not appear to be sustainable, and by 1989 the fruit yield response to applied N continued to the highest N level tested. In that year, the leaf N concentration associated with maximum yield was 1.8 mmol g^–1, the same as that recorded throughout the experiment for the vines planted at high density. In the last two seasons of the experiment, leaf necrosis developed extensively on vines receiving less than the highest rate of N. This necrosis appeared to be premature senescence resulting from N deficiency. Leaf chloride (Cl) concentrations increased significantly with increasing severity of N deficiency, but were never more than those associated with Cl toxicity. While N supply significantly affected fruit firmness immediately post-harvest, there were no significant effects on fruit firmness after 12–20 weeks storage.     

Optimized paired‐sgRNA/Cas9 cloning and expression cassette triggers high‐efficiency multiplex genome editing in kiwifruit

Kiwifruit is an important fruit crop; however, technologies for its functional genomic and molecular improvement are limited. The clustered regulatory interspaced short palindromic repeats (CRISPR)/CRISPR‐associated protein (Cas) system has been successfully applied to genetic improvement in many crops, but its editing capability is variable depending on the different combinations of the synthetic guide RNA (sgRNA) and Cas9 protein expression devices. Optimizing conditions for its use within a particular species is therefore needed to achieve highly efficient genome editing. In this study, we developed a new cloning strategy for generating paired‐sgRNA/Cas9 vectors containing four sgRNAs targeting the kiwifruit phytoene desaturase gene (AcPDS). Comparing to the previous method of paired‐sgRNA cloning, our strategy only requires the synthesis of two gRNA‐containing primers which largely reduces the cost. We further compared efficiencies of paired‐sgRNA/Cas9 vectors containing different sgRNA expression devices, including both the polycistronic tRNA‐sgRNA cassette (PTG) and the traditional CRISPR expression cassette. We found the mutagenesis frequency of the PTG/Cas9 system was 10‐fold higher than that of the CRISPR/Cas9 system, coinciding with the relative expressions of sgRNAs in two different expression cassettes. In particular, we identified large chromosomal fragment deletions induced by the paired‐sgRNAs of the PTG/Cas9 system. Finally, as expected, we found both systems can successfully induce the albino phenotype of kiwifruit plantlets regenerated from the G418‐resistance callus lines. We conclude that the PTG/Cas9 system is a more powerful system than the traditional CRISPR/Cas9 system for kiwifruit genome editing, which provides valuable clues for optimizing CRISPR/Cas9 editing system in other plants.     

Microcallus formation from leaf mesophyll protoplasts in the genus Actinidia Lindl

Summary Microcallus (more than 60 cells) formation was obtained from leaf mesophyll protoplasts of 6 species and varieties in the genus Actinidia Lindl. (kiwifruit). The best results were achieved by using liquid over agarose culture for A. arguta var. arguta, liquid and agarose disc type culture for A. arguta var. purpurea, agarose disc type culture for A. arguta cv. Issaï and A. deliciosa and liquid agarose bead type- and disc type culture for A. kolomikta and A. polygama. Several factors influencing purification, browning, survival and sustained division of the protoplasts are briefly discussed.Abbreviations BAP benzylaminopurine - CPW cell and protoplast washing solution - 2,4-D dichlorophenoxyacetic acid - GA3 gibberellic acid - IAA indole-3-acetic acid - MES 2-(N-morpholino) ethanesulfonic acid - NAA 1-naphthaleneacetic acid - PVP polyvinylpyrrolidone - BT agarose bead type culture - DT agarose disc type culture     

Oryzalin combined with adventitious regeneration for an efficient chromosome doubling of trihaploid kiwifruit

Summary Chromosome doubling of one parthenogenetic trihaploid from cultivar Hayward ofActinidia deliciosa was investigated. Two antimitotic agents, colchicine and oryzalin, appliedin vitro on shoots and leaves at different concentrations were compared with regard to their efficiency. Survival and regeneration rates were determined and ploidy level of regenerated plantlets was evaluated by flow cytometry. Differences were observed between the two antimitotic agents depending on whether shoots or leaves were treated. Hexaploid plantlets were obtained with highest efficiency by adventitious regeneration from leaves treated by oryzalin at 5 M, constituting an original and promising result which was corroborated for another trihaploid clone. Dodecaploid plantlets were also induced but only from oryzalin treated leaves. On the other hand, colchicine applied to leaves was very phytotoxic. This study demonstrates that oryzalin combined with adventitious regeneration is particularly efficient to induce chromosome doubling of trihaploid kiwifruit.Abbreviations MS Murashige and Skoog - IBA indole-3-butyric acid - DMSO dimethylsulfoxide - PBS phosphate buffer salin - DTT dithiothreitol     

猕猴桃溃疡病抗性育种研究进展

猕猴桃细菌性溃疡病是一种危害世界猕猴桃生产的毁灭性病害,目前尚未有有效的防治办法。培育抗性品种是保证猕猴桃产业健康发展的重要途径之一,猕猴桃溃疡病抗性育种成为近年来猕猴桃研究的热点。但是,目前大部分猕猴桃种质资源对溃疡病的抗性不明,限制了猕猴桃优异抗性种质资源的发掘和利用。虽然人们发展出了一些猕猴桃溃疡病抗性鉴定和评价方法,但是使用效果并不理想,存在较大的局限性,鉴定的准确性和稳定性还有待提高。该文针对猕猴桃溃疡病抗性育种中的几个方面,如抗性材料的选育(现有品种的抗性、抗性砧木研究和野生抗溃资源等),抗性鉴定和评价技术(大田鉴定、活体或离体鉴定等)及抗性机理研究等进行综述,并针对存在的问题,提出建设性意见。在猕猴桃溃疡病抗性育种过程中,最关键的是要建立一个科学、系统的溃疡病抗性评价体系,以对猕猴桃种质资源进行大规模的抗性普查和评估,在此基础上充分利用种间杂交和工程育种技术加快抗性育种进程,并以此带动猕猴桃溃疡病抗性机理的深入研究和抗病基因的挖掘和利用等,旨在从根本上解决猕猴桃生产中受溃疡病困扰这一关键难题,促进猕猴桃产业绿色、健康和可持续性发展。     

噬菌体鸡尾酒对猕猴桃溃疡病的田间防治效果及叶际细菌群落结构的影响

【背景】噬菌体鸡尾酒可作为一种杀灭猕猴桃溃疡病病原菌(Pseudomonassyringaepv.actinidiae, Psa)的生物制剂,但关于噬菌体鸡尾酒在田间的防治效果和对猕猴桃植株叶际内生细菌群落结构影响的研究依然较少。【目的】探究噬菌体鸡尾酒在田间防控猕猴桃溃疡病的效果,以及噬菌体鸡尾酒对猕猴桃茎内叶际细菌微生态的影响。【方法】使用猕猴桃溃疡病病原菌感染健康植株,对比施用噬菌体鸡尾酒和传统铜制剂后溃疡病的发病情况,利用高通量测序技术分析猕猴桃叶际内生细菌群落结构的变化。【结果】与铜制剂相比,噬菌体鸡尾酒可更有效地控制猕猴桃溃疡病,改变叶际细菌群落的丰富度与多样性,增强群落结构的稳定性,改善群落物种功能基因丰度情况,一定程度使叶际细菌群落恢复至健康状态。【结论】噬菌体鸡尾酒在杀灭病原菌的同时具有良好的微生态调节功能,在猕猴桃溃疡病的生物防治中具有巨大的应用潜力。     

CPPU对美味猕猴桃果实重量、可溶性固形物及糖酸含量的影响

谢花后用20mg/L CPPU浸果,对美味猕猴桃单果重、可溶性固形物及酸含量均有影响。(1)谢花后10d处理一次和谢花后10d及17d处理两次都可使单果重明显增加,且两次处理优于一次处理。(2)经CPPU处理过的果实可溶性固形物含量下降,但酸度无显著影响。(3)CPPU处理的畸形果比率比对照明显上升。     

SOD猕猴桃果汁对体液免疫、血清与红细胞丙二醛水平的影响

目的 研究超氧化物歧化酶（ＳＯＤ）猕猴桃果汁姑降低血清、红细胞丙二醛（ＭＤＡ）含量及提高机体免疫球白水平的作用。方法 测定ＳＯＤ弥猴桃果汁服用前后正常妇女红细胞ＭＤＡ及血清ＭＤＡ、免疫球蛋白ＩｇＧ、ＩｇＡ、ＩｇＭ的含量。结果 ＳＯＤ弥桃果汁服用后,红细胞与血清ＭＤＡ含量显著降低血清,免疫球蛋白ＩｇＧ、ＩｇＡ、ＩｇＭ的含量。结果 ＳＯＤ猕猴桃果取用后,红细胞与血清ＭＤＡ量显著降低,免疫球蛋白ＩｇＧ、