克拉豆的组织培养与快速繁殖

1植物名称克拉豆(Cratylia argentea Benth.)。2材料类别幼嫩种子。3培养条件(1)种子萌发培养基:MS基本培养基;(2)分化培养基:MS 6-BA3mg·L-1(单位下同) NAA0.3;(3)生根培养基:1/2MS NAA1.0     

Embryological observations on Tipuana tipu (Benth.) O. Kuntze (Dalbergieae,Papilionaceae)

Abstract

The anther and ovule development of Tipuana tipu is described. Some embryological aspects previously not reported for the family have been pointed out, i.e. the Basic type of anther wall development; the constant occurrence of several megasporo-cytes undergoing meiosis, the concurrent development of more than one megaspore and finally the coexistence of three or four mature gametophytes in the same ovule.     

Karyotypic studies on species of Phaseolus (Fabaceae: Phaseolinae)

Ten karyotypes of Phaseolus were analyzed. Possible directions of karyological evolution of those species are suggested. All species studied have karyotypes composed exclusively of metacentric and submetacentric chromosomes, except P. filiformis with all metacentrics, and P. chiapasanus with one pair of subtelocentric chromosomes. Secondary constrictions and satellites are common among species studied. Phaseolus leptostachyus, P. micranthus, and P. macvaughii are considered as aneuploids with 2n= 20, whereas the rest of the species have 2n= 22.     

基于小RNA测序的风轮菜microRNA及其靶基因分析

本研究以风轮菜(Clinopodium chinense (Benth.) O. Kuntze)为材料,采用BGISEQ-500测序平台对风轮菜根、茎和叶的小RNA进行转录组测序,并对其黄酮类物质合成途径中参与调控的microRNA(miRNA)及其靶基因进行了分析。结果显示,鉴定出的保守miRNA有86个,属于26个家族,新发现miRNA 8个,筛选出风轮菜黄酮类物质合成途径中调控3个关键酶的候选miRNA(novel＿mir3、miR167d-5p、miR396h)。通过对靶基因编码的关键酶4-香豆酸辅酶A连接酶进行序列分析和同源建模,发现其具有高度保守的底物结合区域、催化结构域及两个保守的肽基序。     

Phytotoxicity of extracts and fractions of Ouratea spectabilis (Mart. ex Engl.) Engl. (Ochnaceae)

Among the numerous plant species occurring in the Cerrado, Ouratea spectabilis stands out because of the lack of species that grow beneath its canopy. Therefore, this study aimed to evaluate the phytotoxic potential of different extracts and fractions of the hydroethanolic extract from leaves of O. spectabilis through laboratory bioassays of the pre-and post-emergence of seeds of Lactuca sativa L., determination of the mitotic index in root cells of Allium cepa L., antioxidant activity and phytochemical screening of different classes present in extracts and ethyl acetate fractions. It was possible to verify that different extracts and ethyl acetate fractions of O. spectabilis interfered with germination rates, as reduced germination was observed when compared with the control. Similarly, growth and development was affected in lettuce seedlings, as shown by the reduced length of primary roots and hypocotyls compared with the control. In addition, the mitotic index was reduced in treated groups compared with the negative control. HPLC-PAD analysis for both the hydroethanolic extract and its ethyl acetate fraction, showed a predominance of flavonoid compounds belonging to the groups of isoflavones and catechins in ethyl acetate fractions of hydroethanolic extracts. Thus, it was concluded that this species synthesizes phytotoxic compounds capable of interfering in the stabilization and development of other species.     

New flavans isolated from the leaves and stems of Cratylia mollis (Leguminosae)

Cratylia mollis Mart ex. Benth is a species belonging to the Leguminosae family that exists throughout South America, and it is one of the most abundant plants in northeastern Brazil, especially in the semiarid region. This plant is popularly known as “camaratu” and “camaratuba”, and the leaves and stems of this species are used as a substitute for cattle's alimentation during the dry season. The chemical investigation of the methanolic extract from leaves and stems of C. mollis led to the isolation of new flavans named 4,2′,3′-trihydroxy-4′-methoxy-6,7-(methylenedioxy) isoflavan, 7,2′-dihydroxy-6-methoxyflavan, 7,3′-dihydroxy-6,2′-dimethoxyflavan, 7-hydroxy-6-methoxyflavan, 2′-hydroxy-6,7-(methylenedioxy) flavan, 2R*,3S*-7,2′-dihydroxy-6-methoxy-flavan-3-ol, and 2R*,3S*-7,3-dihydroxy-6,2′-dimethoxyflavan-3-ol and an unusual flavan (11H-benzofuro[3,2-b][1] benzopyran-2-methoxy,3-hydroxy,5a,10a-dihydro) named (3R*,2R*)-3-O-2′-7-hydroxy-6-methoxyflavan. The structures of the new compounds were determined using spectroscopic methods.     

Micropropagation of tea (Camellia sinensis (L.) O. Kuntze) using Thidiazuron

The effect of thidiazuron (TDZ) on the micropropagation of Camellia sinensis (China hybrid) was compared with that of benzylaminopurine (BAP) using nodal segments from in vitro raised seedlings. Extremely low concentrations of TDZ (1pM–100nM) alone were effective in inducing shoot bud proliferation and maintaining high rates of shoot multiplication on hormone-free media. On the other hand, higher concentrations of BAP (1–10M) and its continued presence were required to initiate and sustain shoot proliferation. While wider ranges of BAP combined favourably with auxins like NAA or IBA, only specific combinations of TDZ and NAA were effective for shoot proliferation. TDZ treated explants yielded healthy shoots, with sturdy leaves, even during the initial stages of growth, whereas, the effect of BAP was cumulative over subcultures in attaining a high proliferative rate.     

Pollination and breeding system of Couepia uiti (Mart. and Zucc.) Benth (Chrysobalanaceae) in the Pantanal da Nhecolandia.

The pollination biology and breeding system of Couepia uiti was studied. In this species, flowers opened at 06:00 AM anthesis, and nectar production began at around 0800 h, reached a maximum volume from 09:30 AM to 10:30 AM, and decreased thereafter. The nectar sugar concentration increased continuously, but showed an abrupt increase from 10:00 AM to 12:00 AM. Pollen release occurred at about 09:30 AM and was quickly collected. The stigmas became receptive at around 12:00 AM. The pollinators of C. uiti included the bees Apis mellifera, Xylocopa sp. and Bombus sp., and three species of wasps. This conclusion was based on the observation that these hymenopterans had C. uiti pollen on their bodies, visited the receptive flowers, and touched the anthers and stigmas, thereby promoting pollination. Of these floral visitors, A. mellifera was considered to be the most efficient pollinator. However, mixed pollination also occurred. The number of C. uiti flowers visited in the morning (n = 52) was three times smaller than in the afternoon (n = 62), and the species richness of floral visitors was also bigger in the afternoon (eight in the afternoon versus five in the morning). This finding indicated that these floral visitors preferred to exploit nectar rather than pollen. Controlled pollination experiments showed that C. uiti was a self-incompatible species that produced fruits only by cross-pollination. Treatments such as agamospermy and spontaneous and self-pollinations did not produce fruits.     

Boron re-translocation in tea (Camellia sinensis (L.) O. Kuntze) plants

Boron (B) re-translocation is an important factor determining tolerance to B deficiency in plants. In this work growth, B content of leaves with different ages, B partitioning between soluble and cell wall (CW) fractions, and B re-translocation were investigated in tea (Camellia sinensis (L.) O. Kuntze) plants grown hydroponically without (<2.5 μM) and with adequate (46 μM) B supply. Under B deficiency, the proportion of CW bound B increased in the old leaves but decreased in roots. Contrastingly, the proportion of CW bound B was not influenced by B supply in the young leaves. A continuous reduction of B content was observed in all fully expanded leaves as well as in roots of low B plants. Taken together, these results revealed considerable re-translocation of B from mature to growing leaves. Leaf extract and phloem exudate samples were analyzed and sucrose, glucose, and fructose were detected while xylitol, sorbitol, mannitol, maltose, galactose, cellobiose or rafinose were not found in these samples. In the leaf extracts, concentration of sucrose increased under B deficiency conditions, concentration of glucose decreased, while that of fructose remained unchanged. Our results provide circumstantial evidence for a considerable re-translocation of B in tea plants despite lacking polyol compounds.     

In vitro clonal propagation of tea (Camellia sinensis (L.) O. Kuntze)

A system for in vitro clonal propagation has been developed in tea plants. Shoots obtained from primary explants were induced from terminal buds and axillary buds of mature field-grown plants. Cultures were initiated from both types of explants on Murashige and Skoog (MS) medium supplemented with 10% coconut milk (CM), 200 mg l^-1 of yeast extract (YE), 1.4 M indoleacetic acid (IAA) and 17.8 M benzyladenine (BA). The shoot tips were multiplied on 1/2 strength MS medium containing 10% CM, 2.9 M IAA and 17.8 M BA. The larger shoots were separated after multiplication and rooted on 1/2 MS medium supplemented with 11.4 M ascorbic acid and 34.5 M indolebutyric acid (IBA). A pretreatment of the plants with an aqueous solution of 493 M IBA greatly increased the frequency of rooting. More than 60% of the rooted plants have been transferred to soil successfully.Abbreviations BA benzyladenine - IAA indoleacetic acid - IBA indolebutyric acid - YE yeast extract - CM coconut milk - MS Murashige and Skoog medium (1962)     

The Role of the Lens in Seed Imbibition and Seedling Vigour of Sesbania punicea (Cav.) Benth. (Leguminosae: Papilionoideae)

Seeds of Sesbania punicea (Cav.) Benth. are largely impermeable,and the natural, low, percentage which germinate are permeableat the lens. Scarifying seed in acid stimulates germinationand causes various types of damage to the testa, including thelens. Placing seeds in boiling water stimulates germinationby rendering the seeds permeable at the lens. Seeds were mechanicallyscarified at precise sites on the testa to assess influenceof the site and degree of damage on germination. Results indicatethat both the site of initial water entry and the rate of waterentry are important in determining seed and seedling vigour.Damaging the lens is more effective in stimulating germinationand reducing seedling abnormalities than complete excision ofthe lens. This effect can be reversed if seeds from which thelens was excised are allowed to imbibe gradually in inert osmotica.The leguminous lens thus acts as the site of initial water entryand regulates the rate of water uptake, thereby increasing seedand seedling vigour. Sesbania punicea (Cav.) Benth., hardseededness, lens, seed germination, imbibition damage, seedling vigour     

弥散山蚂蝗与大叶拿身草(豆科:蝶形花亚科)的分类学关系

弥散山蚂蝗Desmodium diffusum(新拟)不同于大叶拿身草D. laxiflorum,然而前者在中国却不被承认。弥散山蚂蝗广布于中国,与大叶拿身草相较更为普遍,而大叶拿身草仅分布于中国的广东、广西、台湾及云南南部。崔现举等将弥散山蚂蝗置于单序山蚂蝗D. unibotryosum种中。根据国际命名法规,单序山蚂蝗为一非法名称,属弥散山蚂蝗的异名。本文介绍了弥散山蚂蝗和大叶拿身草的分类历史、种的检索表、种的文献、异名及分布。同时,指定了弥散山蚂蝗的后选模式。     

Pollination biology of Albizia lebbeck (L.) Benth. (Fabaceae: Mimosoideae) with reference to insect floral visitors

Indian siris, Albizia lebbeck (L.) Benth. (Fabaceae: Mimosoideae) has significant importance to human beings for its multipurpose use. Insects play a crucial role in the pollination biology of flowering plants. In the current study, we studied the pollination biology of A. lebbeck with special reference to insect floral visitors. The effectiveness of floral visitors was investigated in term of visitation frequency, visitation rate and pollen load during 2012 and 2013. In the second experiment, effect of pollinators on yield of A. lebbeck was studied in open and cage pollination experiments. Floral visitor fauna of A. lebbeck included eight-bees, two wasps, two flies, and two butterflies species. Among them, Apis dorsata, Apis florea, Amegilla cingulata, and Nomia oxybeloides had maximum abundance ranging from 349–492, 339–428, 291–342 and 235–255 numbers of individuals, respectively during two flowering seasons. A. dorsata had the highest visitation frequency (6.44 ± 0.49–8.78 ± 0.48 visits/flower/5min) followed by Amegilla cingulata (6.03 ± 0.43–7.99 ± 0.33 visits/flower/5min) and A. florea (3.61 ± 0.31–4.44 ± 0.18 visits/flower/5min). A. dorsata, N. oxybeloides, and Amegilla cingulata had the highest visitation rates (18.904 ± 1.53–11.43 ± 1.17 flower visited/min) and pollen load (15333 ± 336.22–19243 ± 648.45 pollen grains). The open pollinated flowers had significantly higher capsule weight (4.97 ± 0.21 g), seed weight (1.04 ± 0.05 g), seed numbers per pod (9.80 ± 0.34) and seed germination percentage (84.0 ± 1.78%) as compared to caged flowers. The results suggested bees especially A. dorsata, N. oxybeloides and Amegilla cingulata could be effective pollinators of A. lebbeck.     

土圞儿属和旋花豆属(豆科)的孢粉学研究

在扫描电子显微镜下,对豆科土儿属(Ap ios)和旋花豆属(Coch lianthus)7个种植物的花粉进行了观察.结果表明,2个属的花粉均为三孔沟,形状为三角球形或球形.土儿属的花粉可划分为2种类型:肉色土儿(A.carnea)的花粉外壁为典型的网状纹饰,其它种的花粉外壁较光滑,具颗粒状或短条纹状纹饰,研究结果不支持土儿属下亚属的划分;肉色土儿花粉外壁的网状纹饰式样可能反映了其在土儿属中处在较进化的位置.土儿属东亚分布种的花粉类型较为多样,原始和进化2种类型并存,表明东亚可能是土儿属的起源地和演化中心.旋花豆属的花粉特征和肉色土儿相似,说明两者之间的亲缘关系可能较近.     

Agrobacterium tumefaciens-mediated transformation of embryogenic tissues of tea (Camellia sinensis (L.) O. Kuntze)

Embryogenic tissues of tea were cocultivated withAgrobacterium tumefaciens LBA4404. The plasmid pBi121, which contains the neomycin phosphotransferase II (nptII) gene providing kanamycin resistance as a selectable marker and the β-glucuronidase (uidA) reporter gene, was used as binary vector. The highest transformation frequency (12 transformants/g fresh weight [FW] of treated embryogenic tissue) was obtained with 5-day-old tissues grown in liquid medium and cocultivated withAgrobacterium for 2 d in the same medium but containing 50 μM acetosyringone. There was improvement in the recovery of kanamycin-resistant tissues when tissues were first grown for 10 d on a medium containing 350 mg/L Timentin to prevent bacterial overgrowth, before application of the selection pressure. Resistant tissues obtained after 6 wk on kanamycin-selection medium showed stableuidA expression. Polymerase chain reaction demonstrated the presence of the transgenes, while Southern hybridization confirmed their integration into the genome. Transgenic plants were regenerated from transformed tissues within 4 mo after coculture.     

Differential leaf traits of a neotropical tree Cariniana legalis (Mart.) Kuntze (Lecythidaceae): comparing saplings and emergent trees

Cariniana legalis is an emergent tree that reaches the upper canopy in Brazilian Semideciduous Forest. Spatial contrasts in microclimatic conditions between the upper canopy and understorey in a forest may affect morpho-physiological leaf traits. In order to test the hypothesis that the upper canopy is more stressful to leaves than a gap environment we compared emergent trees of C. legalis, 28–29 m in height to gap saplings, 6–9 m in height, for the following parameters: leaf area, leaf mass area (LMA the dry weight:leaf area ratio), leaf thickness, leaf anatomical parameters, stomata conductance, and chlorophyll a fluorescence. Leaves from emergent trees had smaller leaf areas but greater LMA compared to saplings. Leaf thickness, palisade layer thickness, and stomatal density were higher for emergent trees than for saplings. The opposite pattern was observed for spongy layer thickness and spongy/palisade ratio. Stomatal conductance was also higher for emergent tree leaves than for sapling leaves, but the magnitude of depression on stomatal conductance near midday was more pronounced in emergent trees. The potential quantum yield of photosystem II, as determined by the F _v/F _m ratio was lower for leaves from saplings. The lower values of stomatal conductance, indicating restriction in CO₂ diffusion into the mesophyll can be related to higher photoinhibition observed in the saplings. Leaves from emergent trees and saplings exhibited similar values for apparent electron transport rates and non-photochemical quenching. Our results suggest that changes in leaf traits could be associated to dry conditions at the upper canopy as well as to the ontogenetic transition between sapling/emergent tree life stages.     

Cloning and expression of embryogenesis-regulating genes in Araucaria angustifolia (Bert.) O. Kuntze (Brazilian Pine)

Angiosperm and gymnosperm plants evolved from a common ancestor about 300 million years ago. Apart from morphological and structural differences in embryogenesis and seed origin, a set of embryogenesis-regulating genes and the molecular mechanisms involved in embryo development seem to have been conserved alike in both taxa. Few studies have covered molecular aspects of embryogenesis in the Brazilian pine, the only economically important native conifer in Brazil. Thus eight embryogenesis-regulating genes, viz., ARGONAUTE 1, CUP-SHAPED COTYLEDON 1, WUSCHEL-related WOX, S-LOCUS LECTIN PROTEIN KINASE, SCARECROW-like, VICILIN 7S, LEAFY COTYLEDON 1, and REVERSIBLE GLYCOSYLATED POLYPEPTIDE 1, were analyzed through semi-quantitative RT-PCR during embryo development and germination. All the eight were found to be differentially expressed in the various developmental stages of zygotic embryos, seeds and seedling tissues. To our knowledge, this is the first report on embryogenesis-regulating gene expression in members of the Araucariaceae family, as well as in plants with recalcitrant seeds.     

金粉蕨配子体发育及其无配子生殖的研究

采用光学显微镜和半薄切片技术对金粉蕨(Onychium siliculosum(Desv.)C.Chr.)的配子体发育及其无配子生殖进行了研究。金粉蕨孢子黄褐色,具三裂缝,孢子萌发时先产生假根,然后长出原叶体细胞,萌发类型为书带蕨型。原叶体细胞经丝状体、片状体发育为原叶体,发育类型为水蕨型。金粉蕨配子体分枝发达,成熟配子体呈丛状。多次培养表明金粉蕨只能产生精子器,不能产生颈卵器,为专性无配子生殖。半薄切片观察表明配子体生长点下方的细胞经分裂产生一群小型细胞,由它们发育为无配子生殖胚。小型细胞先分化出无性胚芽,再分化出胚根。胚芽与胚根,胚与配子体之间由分化出的导管连接。     

Structure and annual biomass production of Nymphoides peltata (Gmel.) O. Kuntze (Menyanthaceae)

In 1980, the monthly changes in biomass and plant surface area, together with aspects of production of Nymphoides peltata (Gmel.) O. Kuntze were studied in a backwater of the river Waal (The Netherlands). Furthermore, the seasonal changes in the vertical stratification of the biomass were studied in concrete tanks. These seasonal changes were studied with the harvest method, while the estimation of the net primary production was based upon biomass data and turnover rates of various plant parts. The data thus obtained are compared with those of other water plants, especially other floating-leaved macrophytes. In 1980, N. peltata reached its peak biomass in August being 372 g AFDW m⁻² (ash-free dry weight). The annual net productivity of Nymphoides was estimated to be 1036 g AFDW m⁻². The leaf blades and their petioles contributed most to the production.