苹果叶片不定梢诱导及起源的细胞组织学观察

本实验获得了苹果“世界一”、“北海道9号”、“千秋”等18个品种叶片的再生不定梢，研究了基因型、基本培养基、激素、暗培养条件等影响苹果叶片不定梢再生的因素，结果表明，基因型是影响叶片不定梢再生的关键因素，对不定梢起源进行细胞组织学探讨，发现苹果叶片不定梢起源多种多样，既有单细胞起源，又有多细胞起源，既有表皮细胞，又有叶肉细胞；既有维管束薄壁组织，又有叶柄基本组织。     

7个牛品种对氧磷酶1基因(PON1)的EcoRV和Alu Ⅰ酶切多态性及其与部分性状的关联分析

应用RFLP技术对7个牛品种进行了对氧磷酶1(paraoxonase 1,PON1)基因的EcoRV和Alu Ⅰ酶切多态性检测;SAS 9.1的GLM过程中的Duncan's检验对不同基因型与部分性状之间进行了关联分析.结果表明,EcoR Ⅴ位点与体重、日增重、眼肌面积和嫩度之间存在显著相关,AG基因型个体与其它基因型相比在日增重和嫩度上存在显著差异(P<0.05),而AA基因型个体相对于其它基因型具有较大的眼肌面积(P<0.05);Alu Ⅰ位点则与体重、净肉重和嫩度之间具有显著的生物学意义(P<0.05),AA型个体相对来说均优于其它基因型个体(P<0.05).品种间的比较表明,除实验牛始重相对趋于一致外,国外肉牛专门化品种在其它方面均优于国内品种.研究结果提示两个位点可尝试作为牛肉多汁性(嫩度)的标记.     

不同基因型苹果的离体再生特性及其对NaN_3的反应

不同基因型苹果叶片培养物再生特性不同，主要表现在再生频率的高低和再生类型的类异，通过培养基中的激素和培养过程的光照调节，可以有效地提高各基因型品种的再生频率。不同基因型苹果叶片培养物对ＮａＮ３诱变处理的反应不同，总体而言，其存活率和再生频率均随ＮａＮ３浓度的提高而下降。不同基因型苹果的适宜ＮａＮ３浓度应有所不同，以３０％存活率为标准，对Ｍ２６、嗄拉，首红，长富２及岩富进行诱变处理的适宜浓度分别为３     

浓缩苹果清汁生产中的色值控制措施研究

为了提高浓缩苹果清汁的初始色值和控制其在储藏过程中的下降速度,根据浓缩苹果清汁生产工艺,对5个苹果品种、5种酶制剂、3种活性炭、6种树脂和储藏温度5个色值关键控制点进行了在线研究.结果表明:选择黄元帅、鸡冠、威锦苹果为制汁原料,采用XXL果胶酶和AG300L淀粉酶对苹果汁进行酶解,选用CA-10S型活性炭吸附脱色,经超滤后再用LSA-900B和LSA0900C之比为1:1进行吸附脱色,最后采用4℃储藏的新工艺,此工艺生产的浓缩苹果清汁的初始色值达到98.8%,储藏6个月色值保持在95%以上,产品符合出口国标准要求.     

4个柱形苹果品种在甘肃的生长习性及生理特性研究

为探讨柱形苹果在甘肃地区栽植的生态适应性和生长习性，分别对陇南市礼县、天水市秦州区和甘谷县等不同生态地区栽植的4个柱形苹果品种的枝类组成、座果率进行了调查比较，同时进行了叶片光合指标以及果实品质测定。结果表明:鲁加1号树体相对较矮小，主枝细长，侧枝相对较多，花量较大，光合能力强，果个大，产量高，可溶性固形物、Vc、总酚含量高，综合品质优于润太1号和舞乐、舞姿，且硬度小，树体抗寒、抗旱能力强。综合不同柱形苹果品种的各个指标，认为柱形苹果品种鲁加1号更适宜在甘肃地区种植。     

苹果砧木新品种中砧1号

铁是植物生长的必需微量元素,缺铁导致苹果发生失绿症,生长结果受到影响.我国苹果主产区恰在缺铁区域范围内,因此,从丰富的苹果种质资源中筛选出铁高效型资源,通过育种手段选育出耐缺铁苹果砧木新品种,是解决苹果生产上因缺铁造成生长发育受影响、产量品质受损的根本性途径.1984年开始,本项目组从苹果属40个苹果树砧木种质资源中筛选铁高效基因型,发现小金海棠在Fe含量极低的条件下仍然能够正常生长,未表现出缺铁失绿症状,为铁高效型.随后,以小金海棠(Malus xiaojinensis Cheng et Jiang)实生苗为基础,建立小金海棠开放授粉杂交后代群体.1990年,从自然实生群体中初选出育种优系.经复选比较试验,选育出苹果无性系砧木中砧1号.中砧1号染色体数为四倍体(2n=4x=68),具有无融合生殖能力,去雄套袋坐果率85％以上.与苹果栽培品种嫁接亲和性好,苗干直立性好,固地性强,半矮化,矮化程度、提早结果作用及丰产能力均与苹果半矮化砧木M7相近.果实风味甜,适口性好,品质极佳.抗苹果早期落叶病和枝干轮纹病,高抗苹果褪绿叶斑病毒(CLSV)、茎痘病毒(SPV)及茎沟槽病毒(SGV)等潜隐病毒.在石灰母质土壤地区用作苹果自根砧木,可有效避免缺铁黄化现象的发生.     

两个超高产小麦品种施钾效应的差异

通过大田试验和室内分析方法研究了潮土上两个超高产小麦品种吸钾能力、钾利用效率 ,初步研究结果表明 :两个超高产小麦品种在吸钾量、施钾效应、钾肥利用率、利用土壤钾素的能力以及钾效率等方面都存在着差异性 ,温麦 6号是一个耐低钾、钾营养高效基因型品种。     

不同基因型糯玉米氮素吸收利用效率的研究 II.氮素积累动态的基因型差异

为了解糯玉米氮素积累进程的变化规律,阐明不同生育时期氮素积累的基因型差异及其对产量形成的作用,分析了31个糯玉米品种在同一施氮水平下四叶期、拔节期、大口期(12叶期)、开花(吐丝)期、鲜穗采收期和成熟期的植株氮素积累量。结果表明,植株氮素含量随生育进程逐渐下降,植株氮素积累量随生育进程的增加呈不对称的S型曲线变化,可用Richards方程拟合。不同品种各生育时期的氮素含量和积累量均存在显著差异。鲜穗高产品种主要在大口至开花阶段增加了吸氮量;而鲜子粒及成熟子粒高产品种主要在大口至开花,其次在开花至鲜穗采收阶段增加了吸氮量。通径分析表明,氮素积累过程主要影响氮素积累总量的高低,而对氮素利用效率影响较小。氮素积累过程S型曲线的Richards方程特征参数品种间差异显著。最大积累速率大、活跃积累期长、快增期的积累速率大和持续时间长对提高品种的氮素吸收总量有利。属于高产、氮素吸收量大、氮素利用效率高的基因型有6个品种,其大口至开花及开花至鲜穗采收阶段的吸氮量平均值分别为1.136和0.554.g/plant,比其它品种分别高24.3%和37.8%;最大积累速率和快增期的积累速率分别为0.068和0.059.g/(d.plant),比其它品种分别高15.8%和15.9%。活跃积累期和快增期的持续时间平均值分别为63.4和29.5.d,比其它品种分别延长了1.9和0.9d。     

利用SSR技术构建多花黑麦草品种指纹图谱

通过构建我国多花黑麦草主栽品种的SSR标记指纹图谱数据库,以实现对多花黑麦草品种的快速、准确鉴定.本研究利用SSR标记,基于多态性高、稳定性强和连锁群分布均匀的原则,从100对多花黑麦草(Lolium multiflorum Lam.)基因组来源的SSR引物中,筛选出12对引物用来构建了21个多花黑麦草品种(系)的指纹图谱.12对SSR引物共扩增出108种多态性基因型,多态性比率达94.15％,每对引物的基因型从5～22种不等,平均每对引物扩增出9种基因型,多态性信息量(PIC)变幅为0.744～0.934,平均为0.843,此12对引物可以在构建多花黑麦草品种DNA指纹数据库作为核心引物推荐使用.21个多花黑麦草品种(系)基因型间遗传相似性系数在0.4956～0.8571之间,UPGMA聚类分析表明,在相似系数0.69处可将全部材料分为3大类.7对引物在9个品种上具有唯一特征带,采用15-08C单对引物即可将21个多花黑麦草品种完全区分开,该对引物不仅多态性高(PIC=0.934),且具备多个品种的特征谱带.基于该引物建立了供试品种指纹图谱标准模式图,每个品种具有唯一的指纹图谱(带型),为牧草品种的审定和保护以及选配优良杂交组合培育新品种提供了重要的理论依据.     

蜡梅非特异性脂转移蛋白基因nsLTP家族3个成员的分子特征及非生物胁迫应答分析

为了探索脂转移蛋白在蜡梅(Chimonanthus praecox)应对非生物胁迫过程中的可能功能,在构建蜡梅花cDNA文库及EST分析的基础上,通过对文库cDNA克隆全长测序,得到了3个蜡梅非特异性脂转移蛋白的基因(nsLTPs),分别命名为CpLTPI.1、CpLTPI.2和CpLTPI.3,GenBank登录号为FJ889521、FJ904082和FJ904083.它们的cDNA全长分别为611,1016和656 bp,ORF分别为360,360和351 bp,5'和3'端的非翻译区的长度不等,存在的调控基序有所不同.3个基因的编码蛋白均具有植物nsLTP的典型结构,即4对二硫键,4个α-螺旋,有1个可结合和容纳脂肪酸分子的类似口袋状的疏水结构,分子量约为9kD,为nsLTPI类.定量RT-PCR分析和比较蜡梅叶片中3个基因对非生物胁迫的应答情况,结果表明,3个nsLTP基因在蜡梅幼苗中的表达受干旱、ABA、低温和NaCl处理的影响程度不同,表明这些基因可能在蜡梅幼苗的水分平衡、耐受低温、离子代谢等过程中发挥着不同的作用.     

Evaluation of genetic identity and variation of local apple cultivars (<Emphasis Type="Italic">Malus</Emphasis> × <Emphasis Type="Italic">domestica</Emphasis> Borkh.) from Spain using microsatellite markers

A collection of 114 local cultivars of Malus × domestica Borkh. from the Northwest of Spain and a group of 26 non-native commercial cultivars, were studied in order to determine the level of genetic differentiation between them, to know the population genetic structure of the three main production regions in North Spain, and to detect possible duplications, misidentifications, and intrusions of foreign cultivars into the present day Spanish apple gene pools. Ten primer pairs of microsatellite loci were selected from previous studies on apple. Allelic frequencies were used to estimate the average expected heterozygosity (Hexp) and F-statistics. A Principal Component Analysis and a Cluster Analysis were conducted in order to determine the relationships among cultivars. Considering all populations (Spanish and commercial cultivars) we found 119 alleles with the 10 microsatellites; 10 out of 119 were rare (with frequencies lower than 0.01). Spanish cultivars can be used as references for those alleles for further studies since they are available in Germplasm banks. Triploid cultivars were quite frequent, 39 out of 140, with the highest number occurring in Galicia (29 cultivars). North Spain presented a high variability for apple according to heterozygosity levels. Microsatellites have provided useful information about the singularity of Spanish apple cultivars, revealed six groups of synonymies and five introgressions of commercial cultivars.     

Technological classification of basque cider apple cultivars according to their polyphenolic profiles by pattern recognition analysis

The polyphenolic compositions of 31 Basque cider apple cultivars were determined in pulp, peel, and juice by high-performance liquid chromatography--diode array detection analysis of crude extracts and after thiolysis. Data sets, consisting of individual polyphenol concentrations, total procyanidin content, and the average degree of polymerization of procyanidins, were evaluated by multivariate chemometric techniques, to develop decision rules for classifying apple cultivars technologically into bitter and nonbitter categories. A preliminary study of the data structure was performed by cluster analysis and principal component analysis in each apple material. Bitter apple varieties presented higher contents of flavan-3-ols and/or dihydrochalcones than nonbitter cultivars. Different classification systems for the two categories on the basis of the chemical data were obtained applying several supervised pattern recognition procedures, such as linear discriminant analysis, K-nearest neighbors, soft independent modeling of class analogy, partial least-squares, and multilayer feed forward artificial neural networks. Excellent performance in terms of recognition and prediction abilities for both categories (100% of hits) was achieved in every case (pulp, peel, or juice). Polyphenolic profiles of apple pulp, peel, or juice provide enough information to develop classification criteria for establishing the technological group of apple cultivars (bitter or nonbitter).     

Cultivar and growing region determine the antioxidant polyphenolic concentration and composition of apples grown in New Zealand

Evidence suggests that increasing consumption of fruit and vegetables contributes to improved health and well-being by providing protection from diseases including various cancers and cardiovascular disease. Although there is uncertainty about which components generate this effect, an attractive hypothesis is that the antioxidants are at least partly responsible. We measured the polyphenolic concentrations in 10 different apple cultivars grown commercially in New Zealand, each sourced from three different geographic regions. Our results showed that the concentration of polyphenolics varied among the apple cultivars, with Pacific Queen containing 2.7 times the amount of polyphenolics found in Cox's Orange. Furthermore, there were significant differences in polyphenolic concentrations in fruit from different regions for some cultivars but not for others. We also measured the polyphenolic concentrations in apple skin and flesh and found that on average 46% of the polyphenolics in whole apples were in the skin. Essentially all of the flavonols (quercetin derivatives) were present in the skin. To maximize the intake of apple polyphenols, it is necessary to consume apples of cultivars with high polyphenolic concentrations such as Pacific Queen and include the skin. Our results also showed that there is potential for promoting apple fruit from specific geographical regions because they contained elevated concentrations of antioxidant polyphenolic compounds.     

Identification of unknown apple (<Emphasis Type="Italic">Malus</Emphasis> × <Emphasis Type="Italic">domestica</Emphasis>) cultivars demonstrates the impact of local breeding program on cultivar diversity

Apple (Malus?×?domestica Borkh.) trees, either abandoned or cared for, are common on the North American landscape. These trees can live for decades, and therefore represent a record of large- and small-scale agricultural practices through time. Here, we assessed the genetic diversity and identity of 330 unknown apple trees in northern Minnesota with 9 simple sequence repeat (SSR) markers. The unknown (not identified by cultivar name) trees were compared to >?1000 named cultivars in the U.S. Department of Agriculture Plant Genetic Resources Unit Malus collection and also to each other to identify repeated genotypes. Overall, the 330 unknown trees had high diversity (average H_e?=?0.75), and consisted of 264 unique genotypes. A total of 76 of the unknown trees were matched to 20 different named cultivars, and these cultivars were mainly derived from either the local breeding program at the University of Minnesota, or were Russian cultivars imported for horticulture in the northern US. This study demonstrates the importance of local breeding programs, and also the challenges associated with identifying clones in a genetically diverse crop like apple.     

Polyphenolic profiles of Basque cider apple cultivars and their technological properties

The polyphenolic compositions of 31 Basque cider apple cultivars were determined in pulp, peel, and juice by high-performance liquid chromatography with diode array detection analysis of crude extracts and after thiolysis. Total polyphenols are distributed in a wide concentration range depending on the cultivar. Procyanidins are the class of polyphenols that present major concentrations in apple. Their average degrees of polymerization range from 4 to 8 depending on the cultivar. Apple cultivars were technologically classified into bitter and nonbitter categories using different classification systems obtained by applying several pattern recognition techniques, such as principal component analysis, K-nearest neighbors, soft independent modeling of class analogy, partial least-squares, and multilayer feed-forward-artificial neural networks, to apple pulp, peel, or juice data (individual polyphenol concentrations, total procyanidin content, and the average degree of polymerization of procyanidins). Bitter apple cultivars present higher contents of flavan-3-ols and/or dihydrochalcones than nonbitter cultivars. Detailed knowledge of the polyphenolic profile of each apple cultivar affords information about their susceptibility to oxidation, their sensory properties (bitterness, astringency), and their possible influence on the characteristics and quality of the final product (juice, cider) when apples are processed.     

Specific detection of potentially allergenic peach and apple in foods using polymerase chain reaction

Two PCR methods were developed for specific detection of the trnS-trnG intergenic spacer region of Prunus persica (peach) and the internal transcribed spacer region of Malus domestica (apple). The peach PCR amplified a target-size product from the DNA of 6 P. persica cultivars including 2 nectarine and 1 flat peach cultivar, but not from those of 36 nontarget species including 6 Prunus and 5 other Rosaceae species. The apple PCR amplified a target-size product from the DNA of 5 M. domestica cultivars, but not from those of 41 nontarget species including 7 Maloideae and 9 other Rosaceae species. Both methods detected the target DNA from strawberry jam and cookies spiked with peach and apple at a level equivalent to about 10 μg of total soluble proteins of peach or apple per gram of incurred food. The specificity and sensitivity were considered to be sufficient for the detection of trace amounts of peach or apple contamination in processed foods.     

Diversity of the European indigenous wild apple (Malus sylvestris (L.) Mill.) in the East Ore Mountains (Osterzgebirge), Germany: II. Genetic characterization

In the present study genetic diversity and hybridization with cultivars were investigated in a population of the endangered European wild apple species Malus sylvestris (L.) Mill. with the aim to establish a basis for the implementation of conservation activities and to ensure its long-term preservation. A total of 284 putative M. sylvestris trees located in the East Ore Mountains were investigated along with a standard set of reference apple genotypes proposed by the European Cooperative Program for Plant Genetic Resources (ECPGR) and 13 old apple cultivars often cultivated in Saxony. The genetic analysis was performed using 12 microsatellite markers also recommend by the ECPGR. To differentiate ‘true type’ M. sylvestris individuals, hybrids and apple cultivars (Malus × domestica Borkh.) a model-based cluster analysis was performed using STRUCTURE. Two clusters were identified consisting of M. sylvestris and M. × domestica genotypes. About 40 % of the putative M. sylvestris showed an admixture of the species-specific allele frequencies and were defined as hybrids. The genetic diversity of the ‘true type’ M. sylvestris population was still high but slightly lower than in the apple cultivars especially since some SSR loci were fixed on one or few alleles in the M. sylvestris population. The differentiation parameters between ‘true type’ wild apple and cultivars indicated a clear discrimination between the wild and cultivated apple individuals. This fact confirms our expectation of the existence of ‘true type’ M. sylvestris individuals in the East Ore Mountains and argues for the realization of preservation measures in this area.     

苹果品种用于加工鲜榨汁的适宜性评价

为科学评价苹果品种的鲜榨汁加工适宜性,以122个单果质量在100 g以上的品种为对象,运用相关分析、因子分析、概率分级、层次分析、K-均值聚类、判别分析等方法建立苹果品种鲜榨汁加工适宜性评价技术。结果表明：果实与鲜榨汁间可滴定酸含量、可溶性固形物含量、可溶性糖含量、固酸比和糖酸比5项指标均呈极显著相关,相关系数分别为0.8967、0.9393、0.8413、0.9036和0.9099。果实可溶性固形物含量、固酸比、出汁率、单宁含量等4项指标被确定为苹果品种鲜榨汁加工适宜性评价指标。4项指标均划分为服从或近似服从正态分布的5级,即极低、低、中、高和极高。根据指标权重和指标分级标准,建立了4项指标的评分标准。建立的苹果品种鲜榨汁加工适宜性判别函数有极高的判别准确性,正确判别率达到94.74%（建模样本）和96.55%（检验样本）。筛选出的58个优良品种中,红富士、乔纳金、津轻等43个品种适于加工鲜榨汁,澳洲青苹、红玉、金冠等15个品种极适于加工鲜榨汁。     

Characterization and quantitation of low and high molecular weight phenolic compounds in apple seeds

The phenolic constituents of seeds of 12 different apple cultivars were fractionated by sequential extraction with aqueous acetone (30:70, v/v) and ethyl acetate after hexane extraction of the lipids. Low molecular weight phenolic compounds were individually quantitated by RP-HPLC-DAD. The contents of extractable and nonextractable procyanidins were determined by applying RP-HPLC following thiolysis and n-butanol/HCl hydrolysis, respectively. As expected, the results revealed marked differences of the ethyl acetate extracts, aqueous acetone extracts, and insoluble residues with regard to contents and mean degrees of polymerization of procyanidins. Total phenolic contents in the defatted apple seed residues ranged between 18.4 and 99.8 mg/g. Phloridzin was the most abundant phenolic compound, representing 79-92% of monomeric polyphenols. Yields of phenolic compounds significantly differed among the cultivars under study, with seeds of cider apples generally being richer in phloridzin and catechins than seeds of dessert apple cultivars. This is the first study presenting comprehensive data on the contents of phenolic compounds in apple seeds comprising extractable and nonextractable procyanidins. Furthermore, the present work points out a strategy for the sustainable and complete exploitation of apple seeds as valuable agro-industrial byproducts, in particular as a rich source of phloridzin and antioxidant flavanols.     

苹果果实中毒死蜱残留的品种间差异及套袋对毒死蜱残留的影响

为探讨苹果果实中毒死蜱残留的品种间差异及套袋对其残留的影响作用,采用气相色谱法（GC-FPD）,研究了不同品种苹果果实中毒死蜱的残留动态以及套袋对苹果果实不同部位中毒死蜱残留的影响。结果表明,毒死蜱在苹果果实中的残留量存在着明显的品种差异,其中红富士属于高农药残留的品种,而嘎拉、红将军和83-1-70-3则属于低农药残留的品种。毒死蜱在苹果果实不同部位中的残留量表现出明显差异,果皮是毒死蜱残留的主要部位,其次是全果,果肉中的残留最少。套袋明显减少了毒死蜱在苹果果实中的残留量,不论处理浓度和取样时间如何,套袋苹果果实中毒死蜱的残留量比不套袋至少减少1/3。