Effects of Salt Stress on Anatomical Structure of Leaves of Malus sieversii and Malus robusta

[Objective] This study was conducted to compare leaf anatomical structure of Malus sieversii and M. robusta under different salt concentration levels to determine their tolerance to salt and thus to provide rootstock materials for apple production in salinized soil in Southern Xinjiang. [Method] The experiment was conducted with M. sieversii and M. robusta as test materials. Salt stress was simulated using 8 g/L of NaCI solution, and Hoagland nutrient solution was used instead of NaCI solution as control group （CK）. Samples were collected on the 20^th d of treatment, sliced through paraffin processing. The prepared paraffin sections of M. sieversii and M. robusta were then observed under a light microscope for anatomical structures of leaf, upper epidermis, lower epidermis, palisade tissue and spongy tissue. [Result] Compared with the control, the leaf, upper epidermis, lower epidermis and spongy tissue of M. sieversii and M. robusta under salt stress were thickened at different degrees, while the thickness of the palisade tissue was decreased. Moreover, high salt concentration caused severer damage to the cell structure of M. sieversii than to that of M. robusta, as M. robusta cells maintained better structural integrity. [Conclusion] M. robusta has higher adaptability to salt stress than M. sieversii.

The Growth-Promoting Mechanism of Brevibacillus laterosporus AMCC100017 on Apple Rootstock Malus robusta

Plant growth-promoting rhizobacteria(PGPR) located in rhizobacteria soil are beneficial to plant growth and development. A PGPR strain AMCC100017 of Brevibacillus laterosporus synthesizes the plant hormone IAA in a tryptophan-dependent manner. In this study, the AMCC100017 strain was used to treat Malus robusta, an excellent natural rootstock for apple production, and assess its ability to promote growth. The fresh weight, dry weight, plant height, and lateral root growth of M. robusta were significantly increased with treatment. The presence of the AMCC100017 strain increased IAA content in M. robusta and promoted root secretion of tryptophan. Colonization of the strain in the roots allowed continuous synthesis of IAA and promoted plant growth. In addition, the photosynthetic efficiency in leaves increased after microbial treatment, and the utilization of nitrogen, zinc, iron, copper and magnesium in leaves was increased, which was conducive to photosynthesis.Interestingly, the activities of CAT and SOD, as well as the contents of ROS in plants colonized by AMCC100017 were increased compared to control plants, but the activities of POD and MDA contents were decreased. AMCC100017 strain affected the antioxidant capacity and stress resistance of plants. AMCC100017 strain increased the content of soluble protein and soluble sugar in plants, improved plant metabolic activity and stress resistance. Therefore, AMCC100017 not only increased IAA content and photosynthetic efficiency to promote M. robusta growth, but also affected plant through multiple metabolic pathways.

Effects of Exogenous Manganese (Mn) on Mineral Elements, Polyamines and Antioxidants in Apple Rootstock Malus robusta Rehd.

Manganese(Mn)is one of the essential microelements in all organisms.However,high level of Mn is deleterious to plants.In this study,the effects of exogenous manganese application on mineral element,polyamine(PA)and antioxidant accumulation,as well as polyamine metabolic and antioxidant enzyme activities,were investigated in Malus robusta Rehd.,a widely grown apple rootstock.High level of Mn treatments decreased endogenous Mg,Na,K and Ca contents,but increased Zn content,in a Mn-concentration-dependent manner.Polyamine metabolic assays revealed that,except the content of perchloric acid insoluble bound(PIS-bound)spermine,which increased significantly,the contents of putrescine(Put),spermidine(Spd)and spermine(Spm)all decreased progressively,accompanied with the decreased activities of arginine decarboxylase(ADC,EC 4.1.1.19)and ornithine decarboxylase(ODC,EC 4.1.1.17),and the increased activities of diamine oxidase(DAO,EC 1.4.3.6)and polyamine oxidase(PAO,EC 1.5.3.3).Further antioxidant capacity analyses demonstrated that contents of anthocyanin,non-protein thiols(NPT)and soluble sugar,and the activities of guaiacol peroxidase(POD,EC 1.11.1.7),catalase(CAT,EC 1.11.1.6)and superoxide dismutase(SOD,EC 1.15.1.1),also increased upon different concentrations of Mn treatments.Our results suggest that endogenous ion homeostasis is affected by high level of Mn application,and polyamine and antioxidant metabolism is involved in the responses of M.robusta Rehd.plants to high level of Mn stress.

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.

Genetic Diversity of Volatile Components in Xinjiang Wild Apple (Malus sieversii)

To evaluate genetic relationships using qualitative and/or quantitative differentiation of volatile components in Xinjiang Wild Apple （Malus sieversii （Lebed.） Roem.） and to acquire basic data for the conservation and utilization of the species, aroma components in ripe fruit of M. sieversii obtained from 30 seedlings at Mohe, Gongliu County, Xinjiang Autonomic Region, China, and in ripe fruit of 4 M. purnila cultivars （＇Rails＇, ＇Delicious＇, ＇Golden Delicious＇, and ＇Fuji＇） were analyzed using head space-solid phase microextraction and gas chromatography-mass spectrometry. The results indicated that the values of similarity coefficient concerning volatile types between the two species were in accordance with the evolution of M. pumila cultivars （forms）, and that M. sieversii seedlings showed considerable genetic variations in these aspects： the total content of volatile components, the classes and contents of each compound classes, the segregation ratio, and content of main components. The results showed significant difference among seedlings and wide genetic diversity within the populations. Comparison of the volatile components in M. sieversii with those in M. pumila cultivars showed that the common compounds whose number were larger than five with the contents over 0.04 mg/L simultaneously between M. sieversii and M. pumila cultivars belonged to esters, alcohols, aldehydes or ketones. This suggests fundamental identity in main volatile components of M. sieversii and M. pumila cultivars. The results above sustained the conclusion ＂M. sieversii is probably the ancestor ofM. purnila＂. However, there were 48 compounds present in M. puraila that were not detected in M. sieversii, including 6 character impact components （i.e., propyl acetate, （Z）-3-hexenal, 2-methyl-l-butanol acetate, pentyl acetate, 3-furanmethanol, and benzene acetaldehyde）. This suggested that in the domestication of M. pumila, introgression of other apple species, except for M. sieversii, by interspecies hybridization was possible. There were 177 compounds in total belonging to 11 classes detected in 30 M. sieversii seedlings, including esters, alcohols, ketones, aldehydes, acids, benzene ramifications, terpenes, heterocycles, hydrocarbon derivates, acetals, and lactones. Among them, acetals and lactones were not detected in M. pumila cultivars, 90 compounds were unique to M. sieversii, and 7 components （l-butanol, ethyl hutanoate, 1-hexanol, ethyl hexanoate, 3-octen-1-ol, ethyl octanoate, and damascenone） belonged to character impact odors. Thus, the potential of M. sieversii in ＂utilization conservation＂ is enormous as a rate germplasm on genetic improvement of M. pumila cultivars.

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。

Genetic Structure of Malus sieversii Population from Xinjiang,China,Revealed by SSR Markers

One hundred and nine Malus sieversii accessions from four geographical populations located at Kuerdening in Mohe town, Gongliu County, Jiaowutuohai, in Xinyuan County, Daxigou in Houcheng County of Ily State, and Baerluke Mountain in Yumin County of Tacheng State, Xinjiang Uygur Autonomous Region of China were studied by SSR markers. The purpose of the study was to determine the genetic structure and diversity in these eco-geographical populations with eight pair SSR primers of apple. The results indicated that： an average of 16 bands was detected in the four populations. The percentage of polymorphic bands in Gongliu population （89.06%） was the highest in the four populations. The average Nei＇s gene diversity index was 0.257 for all the loci. Totally, 128 polymorphic loci were detected and the percentage of polymorphic loci （P） was 100%, 88.28%, 84.83%, 87.50%, and 87.12%, respectively, at the species level and Gongliu, Xinyuan, Huocheng, and Yumin population levels. The Nei＇s gene diversity index （H = 0.2619） and Shannon＇s information index （1 = 0.4082） in the species level were higher than in the population level. The Nei＇s gene diversity index and Shannon＇s information index in the four populations were Gongliu 〉 Huocheng 〉 Xinyuan 〉 Yumin. Gongliu population and Xinyuan population were the highest in genetic identity and the closest in genetic distance. Gene flow between the populations was 7.265 based on genetic differentiation coefficient （GST = 0.064）. The UPGMA cluster analysis indicated that the genetic relationships between the Gongliu and Xinyuan population were the closest, and the Yumin population were the farthest with the other three populations. The UPGMA cluster analysis indicated that the four geographical populations located in Gongliu, Xinyuan, Huocheng, and Yumin were relatively independent populations. Concurrently, there was also mild gene exchange between the populations. On the basis of the study of population genetic structure and the highest genetic diversity, Gongliu population should be given a high priority consideration in Malus sieversii population＇s in situ germplasm conservation.

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。

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 。

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。

东北山定子(Malus baccata(L.) Borkh)野生居群表型遗传多样性分析及生态地理分布研究

东北山定子（Malus baccata（L.）Borkh）野生居群集中分布区位于49°11＇-50°40＇N,119°42＇-120°16＇E之间,海拔520-670 m,呈带状分布于大兴安岭西南麓与呼伦贝尔平原交接的河床半湿地,面积约230 km^2,处于原始野生状态。调查423份自然变异类型表明：树龄最大400年生以上,30-50年生为主体占37%;树姿以直立型与开张型为主,垂枝型占14%;株高与冠径均呈正态分布,矮生型占19.8%;果实颜色由黄经红到紫色变异,浓红型占43.8%,黄色与紫色类型分别占4.5%、3.2%;果实酸、酸甜、甜酸共占总群体的72.9%;自然授粉前提下,平均每花序坐果3个的占47.7%,坐果5个的占26.5%,3.5%的成龄树没有果实;叶形与果形有扁圆、圆、卵圆、长椭圆等,但叶形与果形之间无遗传相关性。群体遗传分析参考国内外研究进展,结合分布区的GPS、河流走向、地貌特征、树龄分布、采访记录等推测：东北山定子是山定子组（Malus baccatus Jiang）的地理亚种,集中分布于大兴安岭西南麓的山谷间,海拔800m左右,种子经河水、鸟类等作用,延伊敏河、根河、海拉尔河、雅鲁河等流域自然迁徙扩散至海拉尔的黑羊站、蒙古屯、哈克镇,满洲里黑山头口岸的弘吉拉布等地,在近河的山谷地带逐渐适应繁衍形成二级集中分布区。

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。

新疆野苹果Malus sieversii(Ldb.)Roem花粉形态及其起源演化研究

本文研究了29个新疆野苹果的花粉形态。结果表明,新疆野苹果的花粉为3孔沟、圆球形、近球形、长球形。外壁呈条纹状纹饰,带有穴状穿孔,可分成5类:条大穴纹饰、条小穴纹饰,条穴纹饰、条穴—附属物纹饰及穴短条纹饰。花粉的P/E 值用于探讨苹果属植物的起源与演化具有重要的意义。新疆野苹果以新源县为起源点,向西发展,经巩留、伊宁到达霍城。在苹果属植物中,新疆野苹果可能是苹果亚组中较原始的种类。

塞威氏苹果Malus sieversii (Ldb.) Roem花粉形态的研究

对采自 75个不同居群的塞威氏苹果 (Malussieversii (Ldb .)Roem)的花粉形态进行了观察。结果表明 :塞威氏苹果花粉萌发孔为三孔沟类型 ,外壁具条纹状纹饰 ,条纹间常具小穴或穿孔。根据外壁纹饰的差异将塞威氏苹果花粉划分为六个类型。同时根据花粉资料 。

用MaxEnt模拟湖北海棠(Malus hupehensis)地理分布

采用湖北海棠551个分布数据以及当前生物气候数据,用MaxEnt软件模拟了它的地理分布。结果显示:①潜在分布地区有巴基斯坦北部、印度北部、尼泊尔、不丹、缅甸东北部、越南西北部、广西北部、西藏南部及东南部、日本岛东南侧、朝鲜半岛中南部及沿海地区;河北东北部、山东半岛北部和辽宁南部,其中分布值在0.6～1的地区有不丹、韩国济州岛及附近岛屿和日本及附近岛屿。②分布记录非常少,但分布值已达0.6～1的地区有山西东南部、江苏省南通、四川雅安市汉源和山东崂山。③以分布值在0.6～1的地区来表示现实地理分布格局,自西东向分布为喜马拉雅南部,东南部-横断山-秦岭-巫山-长江中下游山地-武夷山-朝鲜半岛-日本岛。自北向南,北界在太行山南部,南界武夷山西南部。④Jackknife test检测表明干旱季节的平均温度对湖北海棠的分布增益贡献最大,本种自然分布于干旱季节平均气温在-10℃～15℃的地区。

新疆野苹果(Malus sieversii)内生菌的分离与鉴定

采用组织分离法从新疆野苹果茎中分离纯化了内生菌纯菌株,用分子生物学方法对内生菌进行了种属鉴定,扩增了真菌ITS序列,并进行了比较分析。结果显示,从新疆野苹果中分离得到了11株内生菌,分属于2个属3个种,分别为链格孢属(Alternaria sp.)和镰刀菌属(Fusarium sp.),新疆野苹果与枣树、棕榈树和甘蔗等有相同种类的内生菌,结合MS培养基启动培养、PDA法分离可快速得到纯菌株,比PDA多次纯化分离法更简便高效。本文可为抑制新疆野苹果组织培养时产生的内生菌的生长提供参考。