Identification of S-RNase genotype and analysis of its origin and evolutionary patterns in Malus plants

Identification of the S genotype of Malus plants will greatly promote the discovery of new genes,the cultivation and production of apple,the breeding of new varieties,and the origin and evolution of self-incompatibility in Malus plants.In this experiment,88 Malus germplasm resources,such as Aihuahong,Xishuhaitang,and Reguanzi,were used as materials.Seven gene-specific primer combinations were used in the genotype identification.PCR amplification using leaf DNA produced a single S-RNase gene fragment in all materials.The results revealed that 70 of the identified materials obtained a complete S-RNase genotype,while only one S-RNase gene was found in 18 of them.Through homology comparison and analysis,13 S-RNase genotypes were obtained:S_(1)S_(2)(Aihuahong,etc.),S_(1)S_(28)(Xixian Haitang,etc.),S_(1)S_(51)(Hebei Pingdinghaitang),S_(1)S_(3)(Xiangyangcun Daguo,etc.),S_(2)S_(3)(Zhaiyehaitang,etc.),S_(3)S_(51)(Xishan 1),S_(3)S_(28)(Huangselihaerde,etc.),S_(2)S_(28)(Honghaitang,etc.),S_(4)S_(28)(Bo 11),S_(7)S_(28)(Jiuquan Shaguo),S_(10)S_e(Dongchengguan 13),S_(10)S_(21)(Dongxiangjiao)and S_(3)S_(51)(Xiongyue Haitang).Simultaneously,the frequency of the S gene in the tested materials was analyzed.The findings revealed that different S genes had varying frequencies in Malus resources,as well as varying frequencies between intraspecific and interspecific.S_(3) had the highest frequency of 68.18%,followed by S_(1)(42.04%).In addition,the phylogenetic tree and origin evolution analysis revealed that the S gene differentiation was completed prior to the formation of various apple species,that cultivated species also evolved new S genes,and that the S_(50) gene is the oldest S allele in Malus plants.The S_(1),S_(29),and S_(33) genes in apple-cultivated species,on the other hand,may have originated in M.sieversii,M.hupehensis,and M.kansuensis,respectively.In addition to M.sieversii,M.kansuensis and M.sikkimensis may have also played a role in the origin and evolution of some Chinese apples.

Influence of soaking Malus domeri(Bois)Chev.leaves on gut microbiota and metabolites of long-living elderly individuals in Hezhou city,Guangxi,China

To investigate the effects of drinking the soaking of Malus domeri(Bois)Chev.leaves on gut microbiota and metabolites of long-living elderly individuals in Hezhou city,Guangxi,China.It has been reported that longevity is closely related to metabolism and the gut microbiota.The 16S rRNA sequencing and liquid chromatography-mass spectrometry(LC-MS)were used to analysis fecal samples and explore the factors affecting longevity in the region.Interestingly,we discovered,that elderly individuals who had been drinking the soaking of M.domeri(Bois)Chev.leaves for a long time exhibited higher diversity of the gut microbiota than without drinking the soaking,notably.The proportions of Ruminococcaceae and Prevotella were decreased in those who did not drink this soaking.In addition,a total of 106 metabolites were characterized,and the people of long-lived people(>90 years old)and elderly people(<90 years old)who drinking soaking of M.domeri(Bois)Chev.leaves significantly altered the gut microbiota and upregulated levels of haplopine,farnesol,genipic acid,momordicinin,2-hydroxyestrone,hydroxyphenyllactic acid,caffeic acid,sophoraflavanone B,and soyasaponin I.We preliminarily determined that M.domeri(Bois)Chev.leaves consumption may be an important factor affecting longevity in this area.

Malus’ Law Derived from Deterministic Particle Behavior

A polarized beam of energy is usually interpreted as a set of particles, all having the same polarization state. Difference in behavior between the one and the other particle is then explained by a number of counter-intuitive quantum mechanical concepts like probability distribution, superposition, entanglement and quantized spin. Alternatively, I propose that a polarized beam is composed of a set of particles with a cosine distribution of polarization angles within a polarization area. I show that Malus’ law for the intensity of a beam of polarized light can be derived in a straightforward manner from this distribution. I then show that none of the above-mentioned counter-intuitive concepts are necessary to explain particle behavior and that the ontology of particles, passing through a polarizer, can be easily and intuitively understood. I conclude by formulating some questions for follow-up research.

新疆野生苹果(Malus sieversii)的总多酚、总黄酮提取物的抗氧化活性研究

以采集于新疆不同地区的20个野生苹果品种为研究对象,福林酚法测总多酚,分光光度计法测定总黄酮,DPPH、ABTS、还原能力法测定总抗氧化能力,相关系数法分析各个指标之间的相关性。结果显示,不同品种的野生苹果的总多酚、总黄酮及抗氧化活性之间具有显著差异,其中,HDM-23的总多酚含量最高(4714.33μmol没食子酸/100g鲜果),XY-31的总黄酮含量最高(2452.84mg芦丁/100g鲜果)。总多酚、总黄酮及抗氧化活性之间呈极显著(p<0.01)正相关,表明总多酚与总黄酮是野生苹果抗氧化活性的主要贡献物质。

苹果无融合生殖型砧木“76—2”(Malus Sp.)的利用研究初报

“76—2”苹果砧木具有无融合生殖特性,染色体数量为2n=4x=68。乔木,寿命长,根系发达,侧根及须根量尤多。固地性好,无根蘖,管理方便。耐旱、耐涝、耐瘠薄,并可忍耐-45℃的低温。能在pH8.3—8.6的土壤条件下正常生长。经六年的嫁接鉴定,它对故道地区普遍潜带褪绿叶斑病毒(CLSV)、茎痘病毒SPV)、及茎沟槽病毒(SGV)的金冠品种不发生敏感,嫁接部位愈合良好,平整光滑,无大、小脚现象。植株直立生长,半矮化。其矮化程度、提早结果作用及丰产能力均与M7相近。果实风味浓厚,维生素C、可溶性固形物都显著高于八楞海棠或M7为砧的金冠品种,是有望的实生系半矮化砧。

Bonus-Malus模型在银行贷款机制中的应用

将Bonus-Malus模型应用到银行贷款业务,通过调整银行的Bonus-Malus贷款利率来减少贷款欺诈行为。主要是建立借款人的银行个人信用体系,通过借款人上阶段的还款利率和表现决定其下阶段还款利率,这样就提供了一种与完全审计机制不同的奖惩机制。在一些简单假设下可证明Bonus-Malus利率将会消除所有欺诈行为,而非仅仅减少欺诈行为。

关于Malus定律表述的研究

计算了各种检偏器出射最大光强 Ｉｍａｘ 和通过检偏器前的光强 Ｉ０ 的关系，强调了两者的不同，并指出某些教材中在 Ｍａｌｕｓ 定律叙述中的缺陷．

中国新疆野苹果[Malus sieversii(Lebed.) Roem.]群体遗传结构的SSR分析(英文)

以中国新疆伊犁地区的巩留县莫合镇库尔德宁、新源县交吾托海、霍城县大西沟和塔城地区的裕民县巴尔鲁克山4个种下居群的109个新疆野苹果实生株系为材料,利用8对苹果SSR引物进行群体遗传结构的研究。结果表明:8对SSR引物在4个居群中可平均扩增出16条带,其中巩留县居群多态性带数百分比最高为89.06%,各位点平均Nei基因多样度为0.257;4个群体共扩增出128个位点,在种级水平及巩留县、新源县、霍城县和裕民县4个居群水平多态性位点百分比分别为100%、88.28%、84.38%、87.50%、78.12%,种级水平Nei基因多样度(H=0.2619)和香农信息指数(I=0.4082)大于种下居群,4个种下居群Nei基因多样度和香农信息指数比较巩留县>霍城县>新源县>裕民县;巩留县居群和新源县居群遗传一致度最大,遗传距离最近;根据基因分化系数(GST=0.064)值,测得的基因流Nm为7.265。UPGMA聚类分析结果表明,巩留县和新源县居群遗传关系最近,霍城县居群次之,裕民县居群远离其他3个居群,巩留县、新源县、霍城县和裕民县4个居群是相对独立的群体,但同时存在部分基因交流。所有参数分析表明,巩留县遗传多样性最丰富,故在制定原位种质保护计划时应优先考虑巩留县居群。

Aroma Volatile Compound Analysis of SPME Headspace and Extract Samples from Crabapple (Malus sp.) Fruit Using GC-MS

Volatile compounds from the ripened crabapple fruit of six varieties （Red Splendor, Strawberry Parfait, Pink Spire, Radiant, Sparkler, and Flame） were analyzed by the use of the SPME/GC/MS method. The changes in the volatiles between the ripened and upon full maturity fruit states were studied in Red Splendor and Strawberry Parfait. An effort was made to summarize an effective method for searching and identifying new idioplasms containing a particular fruit aroma within Malus. A total of 37 compounds were identified from the sample. The main aroma volatiles of the six varieties of fruit were comprised of 2-hexenal, 3-hexenal, hexanal, 2,4-hexadienal, benzaldehyde, diethyl phtbalate. The main volatile compound of the crabapple fruit was 2-hexenal, but the relative content percentages were different （45.37, 21.98, 33.56, 32.21, 38.60, and 45.88%）. The aroma components accumulated differently as the fruits ripened. The relative content of aldehydes and esters decreased as alcohols increased after the Red Splendor and Strawberry Parfait fruit ripened. For Red Splendor, the main volatile was still 2-hexenal, but the relative content decreased to 42.89%, and the relative content of alcohols increased by 13.86% as aldehydes and esters declined by 12.16 and 7.18%, respectively. For Strawberry Parfait, the main volatile was changed to cyclohexanol, and the relative content increased to 46.43%, while the relative content of alcohols increased by 49.03% as aldehydes and esters declined by 23.74 and 9.34%, respectively.

新疆野苹果(Malus sieversii)超低温保存及其植株再生

以新疆野苹果(Malus sieversii(Lebed.)M.Roem.)无菌试管苗为试材,对其离体茎尖玻璃化超低温保存的影响因素进行研究。结果表明,新疆野苹果茎尖在含有5%二甲基亚砜(DMSO)的0.4mol/L蔗糖培养基上预培养3 d,60%玻璃化溶液(PVS2)中室温装载30m in,PVS20℃下处理40m in,经液氮保存至少24h后,转入继代培养基上再培养,成活率和再生率分别为93.3%和86.7%。再生植株生长和分化正常;同时对再生植株进行SSR标记检测,未发现超低温保存前后的DNA谱带存在差异。

Genetic structure and historical demography of Malus sieversii in the Yili Valley and the western mountains of the Junggar Basin, Xinjiang, China

Malus sieversii, a wild progenitor of domesticated apple, is distributed in western Xinjiang of China, eastern part of Kazakhstan and Kyrgyzstan in Central Asia. To well understand the genetic structure and the his- torical demography of this important germplasm resource, we sampled 15 populations with 110 individuals of Malus sieversii from the Yili Valley and the western mountains of the Junggar Basin, Xinjiang, and sequenced two nrDNA fragments for these 110 individuals. Meanwhile, we modeled and compared species distributions under the current and the Last Glacial Maximum climatic conditions. The results showed that populations of M. sieversfi from Xinjiang had low levels of genetic diversity and genetic differentiation. During the LGM period, populations of M. sieversfi had lost their northern distributions in the western mountains of the Junggar Basin. M. sieversfi has ex- perienced a demographic expansion from the south of the Yili Valley to the north of the western mountains of the Junggar Basin during the warm interglacial epochs. Due to the high sensibility of M. sieversii to disturbance, we proposed more attention should be paid to the M. sieversfi populations in the western mountains of the Junggar Basin.

Physiological and transcriptomic analyses of roots from Malus sieversii under drought stress

Water deficit is one of the main limiting factors for apple growth and production.Root architecture plays an important role in drought tolerance of plants.However,little is known about the molecular basis of root system in apple trees under drought.In this study,we compared root morphology of two widely used rootstocks of apple(R3 and Malus sieversii)under drought.Our results suggested that M.sieversii is more tolerant to drought than R3,since M.sieversii had a higher ratio of root to shoot as well as root hydraulic conductivity under long-term drought conditions.We then performed whole-genome transcriptomic analysis to figure out the molecular basis of root responses in M.sieversii under drought.It was found that genes involved in transcription regulation,signaling or biosynthesis of hormones,and oxidative stress were differentially expressed under drought.Consistent with the gene expression profile,roots of M.sieversii had higher activities of peroxidase(POD)and superoxide dismutase(SOD)under drought,as well as higher content of abscisic acid(ABA)and lower content of auxin.Taken together,our results revealed the physiological and transcriptomic analyses of M.sieversii roots in response to drought.

Nutrient Deficiency Affects Root Architecture of Young Seedlings of Malus hupehensis (Pamp) Rehd. Under Conditions of Artificial Medium Cultivation

What the researchers go in for is to establish models between root architecture （RA） changes and nutrition, mold ideal root architecture of apple trees, improve the nutrient uptake efficiency, and further explore the functional mechanism of nutrient elements during the course of RA construction. The cultivation system of filter paper is utilized to research the effect of nutrient deficiency on the RA of Malus hupehensis （Pamp.） Rehd. There may be eight types of RA. In complete Hogland solution, the main type of RA is ＂lateral roots clustering in the upper and middle regions of primary root＂. With the lack of P, K or Ca, the main type of RA is ＂lateral roots clustering in the upper region primary root＂, and the ＂lateral roots clustering in the upper and middle regions of primary root＂ types of RA decrease. But with shortage of P, the type of lateral roots clustering in the upper and lower regions of primary root increases, and the type of lateral roots clustering in the middle region of primary root decreases, with the types of RA diversified. Under the condition of K deficiency, the type of no lateral root increases and types of lateral roots clustering in the middle region of primary root decrease, and the percentage of such types as ＂no lateral root＂, ＂lateral roots clustering in the upper region of primary root＂, and ＂lateral roots clustering in the upper and middle regions of primary root＂ accounts for 97.9% in all, with the types of RA simplified. With lack of Fe, Mg or Zn, the main type of RA is ＂lateral roots clustering in the upper and middle regions of primary root＂, but the type of lateral roots evenly-distributed on primary root increases. The main type of RA is ＂lateral roots evenlydistributed on primary root＂, under the condition of N deficiency, and the types of RA turn out to be diversified. There exists a close relation between nutrient deficiency and RA changes. Owing to various forms of nutrient deficiency, correspondingly different types of RA have been produced.

Inheritance and Molecular Marker of Resistance to Bot Canker in Malus domestica

Apple bot canker [Botryosphaeria dothidea（Moug.） Ces.et de Not.] is distributed worldwide,resulting in a serious crop loss every year in apple（Malus domestica Borkh.） production.The resistance of each seedling derived from a hybrid population（Jonathan×Golden Delicious） was evaluated by disease index either from natural infection in the field or from inoculation with five isolates of B.dothidea,Ls1,Lw023,Lw048,Mx1,and Zz26.The inheritance of the resistance to bot canker was analyzed via frequency distribution analysis,and microsatellite and AFLP markers linked to the resistance loci were screened.From the binary frequency distribution patterns,it was found that the segregation ratio of resistant/susceptible genotypes infected by pathogen isolates Lw023 and Ls1 was 1：15;and that by Zz26 and Mx1 was 15：1.The variation of resistance was involved in the segregation of two to four alleles of major genes,the resistance was recessive when infected by Lw023 and Ls1,but was dominant when infected with Mx1 and Zz26.A microsatellite maker,CH02a04-450,and two AFLP markers,E-AG/M-GAC-280 and E-AGG/M-CTT-110,were identified,and their map distances to the resistance loci were 5.1,5.1 and 6.2 cM,respectively.The three markers are located in different linkage groups,while CH02a04-450 is on linkage group 2 or 7.E-AG/M-GAC-280 was successfully converted into SCAR159.Finally,CH02a04-450 and SCAR159 were re-examined in inoculated segregation population and presented a good reliability on predicting phenotypes of resistance.

Effects of Cinnamon Acid on Respiratory Rate and Its Related Enzymes Activity in Roots of Seedlings of Malus hupehensis Rehd.

This paper studied the effects of cinnamon acid treatments on the respiratory rate and related enzymes activity in the seedling roots of Malus hupehensis Rehd.It would provide information for understanding the mechanisms of inhibition damage caused by continuous cultivation of apple tree.20 mL of solution containing different concentrations of cinnamon acid was added into container with the tested seedlings.After treatment,the samples were taken periodically and the respiratory rates were measured by OXY-LAB oxygen electrodes under 25°C stable temperature and then the activities of related enzymes were measured.The rates of total respiration and other 2 pathways [tricarboxylic acid cycle （TCA） and pentose phosphate pathway （PPP）] appeared initially an increasing treads and late （on the 3rd d） began to decline.However,they again appeared an increase trend at the end period,on the contrast,the respiratory rate of embden-meyer- hot-parnas （EMP） pathway appeared a stead decline tread but it had a recover on the last day.The respiratory rate of total and 3 pathways were decreased under 125 mg kg-1 （soil）.The dynamic trends of the enzymes activities of pyrophosphate-dependent phosphofructokinase （PFK）,glucose-6-phosphate dehydrogenase （G-6-PDH） and malate dehydrogenase （MDH） showed similarly.In conclusion,treatments of certain concentration of cinnamon acid would inhibit the respiratory rate and related enzymes activity of roots of M.hupehensis Rehd.And the inhibition degrees were positively related with concentration of cinnamon acid treatments.

台湾林檎(Malus doumeri(Bois.)Chev.)地理分布模拟

依据苹果属最新的系统分类学成果,重新鉴定了中国科学院植物研究所等标本馆的标本,选取了243个有价值的分布记录转换成地理坐标数据,用DIVA-GIS绘制了采集标本的地理分布,这种分布式样主要呈西南—华南—华东的横向连续分布和大陆—台湾的间断性分布。用当前生物气候数据(分辨率30 arc s)采用BIO-CLIM模拟的分布与标本记录分布基本情况是吻合的,并发现了一些潜在分布区域。对落在亚洲地区生物气候变量包络内分布点的19个生物气候变量进行了主成分分析,发现雨季降雨量,年温差,最干月降雨量是台湾林檎分布的主导生物气候因子,变种间仅3个主导生物气候因子的生态位有明显间隔,其它因子生态位不规则重叠。比较了用当前气候数据和未来气候数据对台湾林檎模拟的当前、未来分布格局和潜在分布区,结果显示当前和未来的整体分布范围变化较小,只是整个分布区内部各级分布区的范围和分布值有一定变化。依据台湾林檎的生态需求特性和分布区变化的趋势,推测台湾林檎可能起源于云南、广西、老挝和越南交界的区域。

中国新疆野苹果(Malus sieversii(Ldb.)Roem.)种群地理分布格局及其遗传关系研究

利用SSR(简单重复序列)标记技术,对新疆野苹果的5个主要地理居群(128份材料)进行了种内亲缘关系分析。结果表明:(1)5个居群中巩留居群遗传多样性最丰富;(2)群体的AMO-VA分子方差分析得出野苹果居群遗传多样性有92%来自于居群内,8%来自于居群间,说明遗传多样性主要分布在居群内;(3)5个居群遗传距离与基因流呈极显著负相关,与地理距离呈显著正相关,说明野苹果地理分布格局是决定其遗传关系的主要因素;(4)5个居群间的基因流结果推断野苹果种群在历史上曾发生过迁移,根据种群遗传关系以及地理分布格局推断野苹果在冰期后迁移方向从中亚平原由西向东,由南向北迁移,从而形成现代的地理分布。

西府海棠(Malus micromalus) MaMAPK基因的克隆及表达特性

依据高等植物MAPK基因的保守区设计简并引物 ,用RT PCR方法 ,首次从西府海棠幼苗叶片中克隆了促分裂原活化蛋白激酶 (mitogen activatedproteinkinase,MAPK )基因 (MaMAPK )的cDNA全序列。Southern杂交结果表明在西府海棠中存在一个小的MAPK基因家族。 2 0 %PEG处理西府海棠幼苗不同时间后的Northern杂交分析表明 ,该基因在根系和叶片中均有表达 ,随着胁迫时间延长表达量增加 。

Method of Constructing Core Collection for Malus sieversii in Xinjiang,China Using Molecular Markers

The method for constructing core collection ofMalus sieversii based on molecular marker data was proposed. According to 128 SSR allele of 109 M. sieversii, an allele preferred sampling strategy was used to construct M. sieversii core collection, using the UPGMA （unweighted pair-group average method） cluster method according to Nei ＆ Li, SM, and Jaccard genetic distances, by stepwise clustering, and compared with the random sampling strategy. The number of lost allele and t-test of Nei＇s gene diversity and Shannon＇s information index were used to evaluate the representative core collections. The results showed that compared with the random sampling strategy, allele preferred sampling strategy could construct more representative core collections. SM, difference for construction of M. sieversii core collection. Jaccard, and Nei ＆ Li genetic distances had no significant SRAP （sequence-related amplified polymorphism） data and morphological data showed that allele preferred sampling strategy was a good sampling strategy for constructing core collection of M. sieversii. Allele preferred sampling strategy combined with SM, Jaccard, and Nei ＆ Li genetic distances using stepwise clustering was the suitable method for constructing M. sieversii core collection.

新疆野苹果(Malus sieversii(Ledeb.) M.Roem.)植物学性状遗传多样性及相关性分析

为加强野苹果种质资源利用与研究,促进野苹果研究工作,以3年来调查收集的新疆野苹果（赛威士苹果）的129个单株资源为材料,对单果重、果实纵横经、叶片大小等12个数量性状和叶片颜色、叶尖类型等5个质量性状进行遗传多样性及相关性分析。分析结果表明：叶片颜色、叶尖类型、叶姿、叶缘、叶面状态5个质量性状分布频率较集中;单果重、果实纵横经、叶柄长、叶片长宽、可溶性固形物、干周、树高均存在较大变异,变异系数幅度为16%~51%。各性状多样性指数也较大,均值为1.9264,叶片长的多样性指数最小为1.7359;果梗长的多样性指数最大为2.0525。新疆野苹果资源拥有丰富的遗传多样性,在于果实相关性状的多样性指数高,且变异幅度大,表明丰富的遗传多样性是新疆野苹果资源的重要特征。