Five new limonoids isolated from Walsura robusta

Five new toosendanin limonoids with highly oxidative furan ring walsurobustones A-D(1-4),and one new furan ring degraded limonoid walsurobustone E(5)together with one known compound toonapubesic acid B(6)were isolated from the leaves of Walsura robusta.Their structures were elucidated by NMR and MS data.Especially,the absolute configuration of toonapubesic acid B(6)was confirmed by X-ray diffraction study.Compounds 1-6 exhibited good cytotoxicity against the cancer cell lines HL-60,SMMC-7721,A-549,MCF-7,and SW480.

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.

白芨内生菌Ilyonectria robusta的化学成分研究

采用硅胶、Sephadex LH-20、中压制备以及HPLC等色谱分离方法对一株白芨内生真菌Ilyonectria robusta进行化学成分研究,分离得到2个新化合物和4个已知化合物.通过UV、IR、NMR、MS等多种波谱手段并结合文献数据对比,鉴定其结构分别为:butyl((S)-5-oxopyrrolidine-2-carbonyl)-D-leucinate(1)、butyl((S)-5-oxopyrrolidine-2-carbonyl)-D-phenylalaninate(2)、4-hydroxy-17R-methylincistero(3)、polyporenic acid C(4)、perlolyrine(5)和4-(9H-βCarbolin-1-yl)-4-oxobut-2-enoic acid methyl ester(6).以上化合物均首次从该菌中分离得到,化合物1和2为新化合物.采用MTT法检测了化合物1和2对人类乳腺癌细胞MCF-7的细胞毒活性,化合物1对MCF-7具有一定的抑制作用,IC_(50)为36.2µM.

Species diversity and regeneration of old-growth seasonally dry Shorea robusta forests following gap formation

Diversity and regeneration of woody species were investigated in two ecological niches viz. gap and intact vegetation in old-growth seasonally dry Shorea robusta （Gaertn. f.） forests in Nepal. We also related varieties of diversity measures and regeneration at- tributes to gap characteristics. Stem density of tree and shrub components is higher in the gap than in the intact vegetation. Seedling densities of S. robusta and Terminalia alata （B. Heyne ex Roth.） are higher in the gap than in the intact vegetation, while contrary result is observed for T. bellirica （Gaertn. ex Roxb.） and Syzigium cumini （L. Skeels） in term of seedling density. The complement of Simpson index, Evenness index, and species-individual ratio in the seedling layer are lower in the gap than the intact vegetation. Gap size can explain species richness and species establishment rate. Gaps created by multiple tree falls in different years have higher seedling density of S. robusta than gaps created by single and/or multiple tree falls in the same year. In conclusion, gaps maintain species diversity by increasing seedling density, and favor regeneration of Sal forests. In addition to gap size, other gap attributes also affect species diversity and regeneration.

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.

Virulence and mycotoxic effects of Metarhizium anisopliae on Mahogany shoot borer,Hypsipyla robusta(Lepidoptera:Pyralidae)

Developing appropriate control measures for the Mahogany shoot borer, Hypsipyla robusta Moore has become increasingly important due to the severe damaging effect of the pest on the establishment of the saplings of Swietenia mahagoni Jacq （Sapindales： Meliaceae）. Existing management methods are largely limited to silvicultural practices and spraying of chemical insecticides. To identify a potential fungal biocontrol agent, we compared the virulence of six native and two standard ARSEF isolates of Metarhizium anisopliae Metsch. against this pest. The average survival time and conidial yield of IWST-Ma7 was higher （6.2 to 7.3 days and 4.9 to 4.7 x 105 conidia/ml） than the standards. Sig- nificant difference in sporulation on the cadavers between isolates, doses and incubation periods were substantiated for the selection of potential strain. The mycotoxic effects of crude soluble protein extract when in- corporated in the artificial diet, the ARSEF 2596 and ARSEF 3603 showed LDs0 value of 3.7% and 5.6%. However, IWST-Ma7 was highly lethal with significant lowest LDs0 value of 2.6%. The enzyme activity of IWST-Ma7 was highest for chitinase, CDA, protease and lipase viz., 1.90 U/mg, 1.80 U/mg, 0.98 U/mg and 0.80 U/mg respectively. However the enzyme activity of chitinase and Chitin deacetylase assay for all the isolates was significantly higher than protease and lipase activity. The ITS regions （5.8S rDNA and 28S rDNA） of seven isolates of M. anisopliae were amplified using the ITS1 and ITS4 primers which was a unique fragment of approximately 550 bp. Based on ITS regions, phy- logenetic tree have been constructed and the isolates have been grouped in to 5 clades. The virulence and mycotoxic effects of different isolates could rationally be used to employ them for the management of the mahogany borer.

Phylogenetic diversity of bacteria associated with the marine sponge Agelas robusta from South China Sea

It is well known that marine sponges harbor large numbers of bacteria,some of which have been proved to be sponge-specific.The diversity of bacteria in marine sponges distributed along the coast of South China Sea has been previously studied but that of bacteria in sponges inhabiting the open sea has been rarely investigated.In this report,the diversity of bacteria associated with the marine sponge Agelas robusta from a remote coral reef in the South China Sea was documented employing 16S rDNA library construction,amplified ribosomal DNA restriction analysis（ARDRA） and phylogenetic analysis.A total of 32 phylotypes were finally categorized in nine phyla including Cyanobacteria,Proteobacteria,Chloroflexi,Firmicutes,Actinobacteria,Acidobacteria,Planctomycetes,Bacteroidetes and Gemmatimonadetes.The dominant phylum was Proteobacteria whereas the dominant genus was Synechococcus in Cyanobacteria.Some spongespecific bacteria were also found in the bacteria population,but the proportion（5 OTUs/32 OTUs） was much lower than other sponges.This study reveals the phylogenetic diversity of bacteria in A.robusta and confirms the presence of some sponge-specific bacteria in the South China Sea.Understanding the diversity of sponge-associated bacteria in China assists to exploit the bacteria resources for biotechnology.

Euglobal-IIIa, a novel acylphloroglucinol-sesquiterpene derivative from Eucalyptus robusta: absolute structure and cytotoxicity

Euglobal-IIIa (1), a novel acylphloroglucinol-sesquiterpene derivative, and a known analogue, have been isolated from leaves of Eucalyptus robusta. The structures was elucidated by extensive spectroscopic data and by comparison with data reported in literature, while the absolute configuration of 1 was determined by the X-ray diffraction analysis. Compound 1 exhibited comparable cytotoxicity with that of cisplatin against five human cancer cell lines HL-60, SMMC-7721, A-549, MCF-7, and SW480 with IC50 values of 15.7, 15.5, 17.6, 14.3, and 21.8 μM, respectively.

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.

The Effects of Shade Tree Types on Light Variation and Robusta Coffee Production in Vietnam

Vietnam is well-known as the second largest global coffee producer and the largest worldwide exporter of Robusta coffee. However, the Robusta coffee sector in Vietnam is facing many problems, including low quality, high external inputs and water shortages as a result of shade tree eradication. A six-month research project was conducted that focused on effects of shaded tree types on variation of light intensity and aspects of Robusta production. Three shade tree species at different planting densities and shade provision were investigated, including 46 trees of Durian ha–1 (14% shade), 35 trees of Sennaha–1 (17% shade), and 60 trees of Leucaena ha–1 (34% shade), and unshaded site (Open) was used as a control. The study found that light intensity declined 50% with Durian, 58% with Senna and 60% with Leucaena compared with the Open site (2096 μmol?m–2?s–1). Within the coffee canopy, a significant decline in light intensity was observed from the top of the canopy to the bottom. The percentage of light at the middle (90 cm above ground) and bottom (30cm) levels of the coffee canopy was declined by 81% and 88% respectively for the Open site, and 86% and 92% for the Leucaena site. There were no differences in the number of flowers branch–1 tree–1 and fruit set between shaded and unshaded coffee sites. However, a significant difference in first fruit drop was observed.

Derrisisoflavones H-K and One Isoflavan Derivative from Derris robusta

Four hitherto unknown prenylated isoflavonoids,named derrisisoflavones H-K(1-4)and one new isoflavan,namely 6-hydroxyisosativan(5),were isolated from the ethanol extract of Derris robusta.Their structures were elucidated on the basis of extensive spectroscopic studies.To our knowledge,derrisisoflavones J(3)and K(4)are the first examples of hydroxyethylated isoflavonoid.

A study of using grey system theory and artificial neural network on the climbing ability of <i>Buergeria robusta</i>frog

Ecological engineering is an emerging study of integrating both ecology and engineering, concerned with the design, monitoring, and construction of ecosystems. In recent years, the threat to amphibian animals is becoming more and more serious. In particular, the loss of habitats caused by changes to the way land is used by human beings has hit amphibians particularly hard. Amphibians are known to be particularly vulnerable to human activities because they rely on both terrestrial and aquatic habitats for survival. With the increasing development of many areas in recent years, concrete structures are often installed along water bodies in order to increase the safety of local residents. The construction of concrete banks along rivers associated with human development has become a serious problem in Taiwan. Most ecosystems used by amphibians are lakes and stream banks, yet no related design solutions to accommodate the needs of amphibians. The need to develop the relevant design specification considering protecting the amphibian is imperative. Buergeria robusta, an endemic species in Taiwan, is tree frog widely distributed in lowland montane regions. Their breeding season is from April to September. They like to rest on trees or hide at caves during the daytime and move to the stream nearby in dusk for breeding. Males usually emit weak mating call while standing on stones. Sticky eggs are attached to undersides of rocks and stones. Tadpoles are found in slow flowing water of streams [1]. The goal of this study is to improve the understanding of the relationship between the climbing ability and the physical characteristics of amphibians. In this study, we use Artificial Neural Network to simulate the climbing ability of Buergeria robusta. Besides, Grey System Theory is also adopted to improve the performance of Artificial Neural Network. Artificial Neural Network (ANN) is a computing system that uses a large number of artificial neurons imitating natural neural ability to deal with an information network by computing system. The numerical results have show good agreement with the experimental results. The results can serve as a reference for technicians involved in future ecological engineering designs of banks throughout the world.

Pterocarpadiols A–D, Rare 6a,11b-Dihydroxypterocarpans from Derris robusta

Four hitherto unknown 6a,11b-dihydroxypterocarpans,namely pterocarpadiols A–D(1–4),were isolated from the ethanol extract of the twigs and leaves of Derris robusta.Their structures were elucidated on the basis of extensive spectroscopic analysis.Pterocarpadiols A–D are a kind of very rare 6a,11b-dihydroxypterocarpans,and their presence as markers may be helpful in chemotaxonomical classification.

Ability of Non Symbiotic Nitrogen-Fixing Bacteria Isolated from Coffee Plant Rhizosphere and Their Effects on Robusta Coffee Seedlings

Nitrogen is one of the most needed elements by coffee plants. Utilization of biological nitrogen fixation by non symbiotic bacteria offers alternative to reduce the N fertilizer usage. This study was focused to obtain aerobic non symbiotic nitrogen-fixing bacteria from coffee rhizosphere. The application of those bacteria was expected to enhance coffee seedling growth. Sixty four aerobic nitrogen-fixing bacterial isolates were isolated from coffee plants rhizosphere from Jember, East Java using several nitrogen free medium, such as Ashby, malate acid, and fahreus agar. The nitrogen fixation ability of the isolates was determined by measuring their ability in pellicle formation on semi solid medium and ammonium excretion on growth medium. Ab Kws.l, Asm E6s.3.a, Asm Bsl.1, and Asm E6s were the isolates which showed the best performance on nitrogen fixation with excreted ammonium concentration ranged from 129.6 up to 239.8 pM/mg dry weight cell. Acetylene reduction assay was used to detect nitrogenase activity. Ab Kws.1 was the isolate which had the highest nitrogenase activity （7.4 mmol N2 fixed/gram dry weight cell/hour）. Inoculation of the four best isolates onto Robusta coffee seedling positively enhanced the seedling growth in this green house experiment. Based on the results of Becton Dickinson＇s （BD） PhoenixTM Automated Microbiology System biochemical tests, Asm Bls.l isolates has similarities with Achromobacter sp., Asm E6s.l and Asm E6s.3.a had similarities with Stenotrophomonas maltophilia, while Ab Kws. 1 had similarities with Leifsonia aquatica.

The Ability of Phosphate Solubilizing Bacteria Isolated from Coffee Plant Rhizosphere and Their Effects on Robusta Coffee Seedlings

Phosphate solubilizing bacteria isolated from rhizosphere of coffee plants may play an important role in improving phosphate availability for the plants. However, one of the factors influencing the degree of phosphate solubilization by these bacteria is the ability of the microorganisms to utilize phosphate. The objective of this study was to determine the ability of phosphate solubilizing bacteria isolated from coffee plant rhizosphere and their effects on robusta coffee seedling growth. This research was carried out by taking soil samples from Andungsari （Bondowoso District） and Kaliwining （Jember District） coffee plantations, both located in East Java. Liquid medium of Pikovskaya was used for isolation of phosphate solubilizing bacteria from the soil samples. Results of this study showed that 12 phosphate solubilizing bacteria were obtained from this isolation, eight isolates from Andungsari and four isolates from Kaliwining. Selection of those bacteria isolates was based on the qualitative ability in phosphate solubilizing by measuring the clear zone surrounding the colonies and quantitatively by measuring the solubilized phosphate using spectrophotometer. The results showed that four isolates, in the order of PFpKW1, PFpC61, PFsC62a, and PFsB 11, had the highest qualitative ability in solubilizing phosphate, while for the highest quantitative ability the order was PFpKW 1, PFpC61, PFsC62a, and PFsB 11. In a green house study, inoculation of these selected isolates onto Robusta coffee seedlings positively enhanced the coffee seedling growth. Phenotypic test indicated that the four isolates are similar to the genus of Pseudomonas.

采后处理对“Robusta”香蕉货架寿命及品质的影响

目前印度正在不断扩大种植“Robusta”香蕉。这就要求有比较可行的办法延长该品种的货架寿命以适应内外贸易的需要。本研究包括以下三个方面:(i)保鲜剂Waxol和Tal prolong延迟后熟的效果;(ii)聚乙烯袋包装的保鲜效果;(iii)对未处理果应用乙烯吸收剂。

Responses of Apple Rootstock Leaf Structure on Water Stress

[Objective] The aim was to explore, leaf structure characters of Malus rockii and Malus robusta at drought and rewatedng. [Method] The research used Malus rockii and Malus robusta as test materials to conduct water stress treatment by water control and rewatering. [Result] Malus rockii consists of two layers of pal- isade tissue and Malus robusta consist of three layers of columnar cells. Leaf of Malus robusta is thicker, and the thickness of leaf palisade tissue-to-the thickness of spongy tissue ratio is higher. In contrast, leaf stoma of Malus rockii performs more sensitively to water change, and the stoma area after rewatering is 10 times as high as Malus robusta. Besides, after rewatering, SR grows higher, the thickness of leaf palisade tissue-to-the thickness of spongy tissue ratio declines of the two test materials, but the changes of Malus robusta prove more significant. [Conclusion] The two test materials are different in responses to drought and rewatering, and the leaf structure characters of Malus robusta changes obviously than that of Malus rockii during water variation in terms of resisting drought.

Predicting the upper stem diameters and volume of a tropical dominant tree species

Predicting upper stem diameters and individual tree volumes is important for product quantification and can provide important information for the sustainable management of forests of important commercial tree species(Shorea robusta)in Nepal.The aim of this study was to develop a taper equation for S.robusta.Fifty-four trees were selected and felled in the southern low land of Nepal.A mixed effect modelling approach was used to evaluate 17 different taper functions.‘Leave-one-out cross-validation'was used to validate the fitted taper functions.The variable exponent taper function best fitted our data and described more than 99%of the variation in upper stem diameters.Results also showed significant effects of stand density on tree taper.Individual tree volume prediction using the local volume model developed in this study was more accurate compared to the volume predicted through the taper function and existing volume model.The taper function developed in this study provides the benefit of predicting upper stem diameter and can be used for predicting volume to any merchantable height of individual trees.It will have implications in estimates of volume,biomass,and carbon and thus may be a potential supporting tool in carbon trade and revenue generation.

Silvicultural Systems for Restoration of Mahogany in Degraded Landscapes in Africa: Influence of Mixed Rainforest Plantation on Growth and Pest Damage

African mahogany, one of the world’s most valuable timber species is threatened by over-exploitation in natural forests and failure of plantations due to attacks by the shoot borer Hypsipyla robusta. Mixed-species plantations has been reported to be an effective component of integrated pest management of major pest in other crops;but there is very limited empirical data on its use for managing Hypsipyla in mahogany mixed stands in West Africa. The aim of this study was to assess the effect of mixed-species stands as management intervention, on the growth of Khaya grandifoliola and Khaya ivorensis in relation to Hypsipyla robusta attack in a 10 ha experimental plantation in the wet evergreen forest type in Ghana. Khaya grandifoliola recorded faster growth than Khaya ivorensis in this forest type though the later naturally grow in this forest type while the former is introduced from the dry forest. Two years after planting, diameter and height growth were greater in the mixed-species stand than the pure stands for K. grandifoliola and K. ivorensis. Hypsipyla damage was less in the mixed stands of both K. grandifoliola and K. ivorensis compared to the pure stands, with the 20% and 10% Khaya mixed stand recording the lowest attack in both species. It can be recommended that mixed stands of the two Khaya species at 20% or lower Khaya density might be ideal for reducing the levels of Hypsipyla attack in this type of forest.

Fossils of Macaca mulatta from Tianyuan Cave, with Notes on Some Previously Unpublished Specimens of Macaca from Zhoukoudian Area

The recently discovered macaque fossils, including isolated teeth and mandible fragments, from Tianyuan Cave and the juvenile maxilla from Wanglaopu Cave near Zhoukoudian can be identified as Macaca mulatta; they represent the first recognized occurrence of fossil M. mulatta in the far north of China. Some previously unearthed but still unpublished Macaca specimens were also studied, including a cranium and the associated mandible from CKT Loc.13A, a maxilla and a mandible from Loc.1, a mandible from Loc.4, and a maxilla from Loc.20, as well as a mandible with complete toothrow from Zhoukoudian but without locality details, all of them can be referred to M. robusta. The study shows M. robusta is more closely related to M. mulatta than to M. anderssonL Zhoukoudian is the richest area of Macacina fossils in North China, totally 2 genera and 4 species were once recognized from 9 localities, namely, Procynocephalus wimani, M. cf. anderssoni, M. robusta and M. mulatta. The fossil records of Macacina in Zhoukoudian area were almost continuous through the Quaternary Period, and it＇s reasonable to assume that Zhoukoudian used to be an evolutionary center for macaques in North China. Therefore, the macaque population of M. mulatta once existed decades ago around Beijing probably originated from the Pleistocene ancestors in the local area, rather than being human-introduced as previously thought.