Exploring the attenuation mechanisms of Dalbergia odorifera leaves extract on cerebral ischemia-reperfusion based on weighted gene co-expression network analysis

The attenuation function of Dalbergia odorifera leaves on cerebral ischemia-reperfusion(I/R)is little known.The candidate targets for the Chinese herb were extracted from brain tissues through the high-affinity chromatography.The molecular mechanism of D.odorifera leaves on cerebral I/R was investigated.Methods:Serial affinity chromatography based on D.odorifera leaves extract(DLE)affinity matrices were applied to find specific binding proteins in the brain tissues implemented on C57BL/6 mice by intraluminal middle cerebral artery occlusion for 1 h and reperfusion for 24 h.Specific binding proteins were subjected to mass-spectrometry to search for the differentially expressed proteins between control and DLE-affinity matrices.The hub genes were screened based on weighted gene co-expression network analysis(WGCNA).Then,predictive biology and potential experimental verification were performed for the candidate genes.The protective role of DLE in blood-brain barrier damage in cerebral I/R mice was evaluated by the leakage of Evans blue,western blotting,immunohistochemistry,and immunofluorescent staining.Results:952 differentially expressed proteins were classified into seven modules based on WGCNA under soft threshold 6.Based on WGCNA,AKT1,PIK3CA,NOS3,SMAD3,SMAD1,IL6,MAPK1,TGFBR2,TGFBR1,MAPK3,IGF1R,LRG1,mTOR,ROCK1,TGFB1,IL1B,SMAD2,and SMAD518 candidate hub proteins were involved in turquoise module.TGF-β,MAPK,focal adhesion,and adherens junction signaling pathway were associated with candidate hub proteins.Gene ontology analysis demonstrated that candidate hub proteins were related to the TGF-βreceptor signaling pathway,common-partner SMAD protein phosphorylation,etc.DLE could significantly reduce the leakage of Evans blue in mice with cerebral I/R,while attenuating the expression of occludin,claudin-5,and zonula occludens-1.Western blotting demonstrated that regulation of TGF-β/SMAD signaling pathway played an essential role in the protective effect of DLE.Conclusion:Thus,a number of candidate hub proteins were identified based on DLE affinity chromatography through WGCNA.DLE could attenuate the dysfunction of bloodbrain barrier in the TGF-β/SMAD signaling pathway induced by cerebral I/R.

Discussion on Development of Hainan Huanghuali(Dalbergia odorifera T.Chen)

On the morphological characteristics, growth environment and quality, rarity, and wood characteristics and value of Huanghuali, this paper discusses the development ways of Hainan Huanghuali It is suggested that Halnan Huanghuali could be well grown by guiding farmers to make full use of wasteland, slopes, scattered land, roadside and other resources; the landscaping greenbelt of the courtyard area and the residential area could be planned with nature as a main consideration for planting Huanghuali; and landscaping-type Huanghuali planting could make full use of local environment and surrounding local resources, highlighting the personalized landscaping greenbelt, forming good transition between residential area and urban space and creating a natural kindly living environment for the residents, so as to play the best ecological, social and economic benefits.

Dalbergia prazeri Prain(豆科)之考证

在发表Dalbergia prazeri Prain时Prain曾认为其与托叶黄檀(D.stipulacea Roxb.)近缘,只是该种小叶背面疏被微柔毛,花萼被硬毛与托叶黄檀不同,同时又指出其荚果也与后者不同,但是并没有解释其不同点。之后Prain又将其归并入奥氏黄檀(D.oliveri Gamble ex Prain),亦未给出相应的理由。经研究D.prazeri Prain与南岭黄檀(D.assamica Benth.)为同种,而被归并入后者。Prain发表该种时引证Prazer s.n.为模式,但并没有指定主模式。涉及该种的Prazer s.n.的标本共有6份,该文将藏于印度国立标本馆加尔各答馆(CAL),条形码为CAL0000012326(CAL标本号131311)的标本指定为后选模式(lectotype),其余分别藏于加尔各答标本馆的4份及英国皇家植物园邱园标本馆(K)的1份为等后选模式(isolectotype)。

Cardioprotective effects of Salvia miltiorrhiza Bunge and Lignum dalbergiae odoriferae on rat myocardial ischemia/reperfusion injury

Aim Salvia miltiorrhiza Bunge （SM） and lignum dalbergiae odoriferae （DO） are both traditional Chi- nese medicine that have cardioprotective effects. Here, we further examined the combined effects of SM and DO on rat myocardial ischemia/reperfusion injury. The possible mechanism of SM and DO also were elucidated. Methods DO was divided into aqueous extract of lignum dalbergiae odoriferae （DOW） and lignum dalbergiae odoriferae oil （DOO）. Sprague-Dawley rats were randomized to seven groups： sham group, model group, treatment groups inclu- ding SM （10 g · kg^-1）, DOW （5 g · kg^-1）, DOO （0.5 ml · kg^-1）, SM ＋ DOW （10 g · kg^-1 ＋ 5 g · kg^-1）, SM ＋ DOO （ 10 g · kg^-1 ＋ 0. 5 ml · kg^-1）. Rats were pretreated with homologous drug for 7 days and then subjec- ted to 30 rain of ischemia followed by 180 rain of reperfusion. Electrocardiogram （ECG） and heart rate were moni- tored and recorded continuously. At the end of reperfusion, blood samples were collected to determine the serum levels of creatine kinase-MB （CK-MB） and lactate dehydrogenase （LDH）. Hearts were harvested to assess heart- body rate, infarct size and histopathological changes as well. Maximum and minimum effective points were deter- mined by measuring indicators associate with myocardial injury at different time-points of reperfusion （Smin, 15min, 30min, 45rain, 60min, 120min, 180min）. The potential therapeutic mechanism of SM and SM ＋ DOO were carried out by detecting superoxide dismutase （SOD）, malondialdehyde （MDA）, tumor necrosis factor-alpha （TNF-alpha） and interleukin 6 （IL-6）. Results The results showed SM and DO can ameliorate cardiac function respectively, and this cardioprotective effect was further strengthened by their combinations. Among all the combi- nations, SM ＋ DOO showed predominant potential to improve ECG and heart rate, reduce heart-body rate （28.5% ＋ 1.4% , P 〈 0.01 vs model） and myocardial infarct size （ 20.96% ＋ 1.61% , P 〈 0.01 vs model, P 〈 0.05 vs SM） , attenuate histopathological damage, decrease the levels of CK-MB and LDH （P 〈 0.01 vs model, P 〈 0.05 vs SM）. The maximum effective points of SM and SM ＋ DOO were 15min and 30rain respectively, and the minimum effective points of them were 180rain. In reducing serum level of MDA, TNF-alpha, IL-6 and increasing SOD activ- ity, SM ＋ DOO was similar to SM. Conclusion The results of this study indicated that SM ＋ DOO have combined effects that are highly effective than single pretreatment against myocardial ischemie reperfusion injury in rats. The possible mechanism of SM and DO were likely through its anti-oxidant and anti-inflammatory properties, and thus may be an effective and promising medicine for both prophylaxis and treatment of ischemic heart disease.

Effects of mother tree ages, different rooting mediums, light conditions and auxin treatments on rooting behaviour of Dalbergia sissoo branch cuttings

Dalbergia sissoo Roxb. is one of the promising multipurpose tree species of South Asia. Most of the plantations of D. sissoo from seeds are facing severe threats due to the die-back disease, which ultimately causes death of this potential tree-species within a few months. Vegetative propagation could avoid the die-back disease. Thirty mother trees of different age-groups of D. sissoo were selected for evaluating the rooting behaviour of branch cuttings from D. sissoo as influenced by auxins （IAA or IBA at 100, 200, 500 mg·L^-1）, ages of mother trees （10, 4 and 2 years old） and different environment conditions, i.e., different mediums （soil and sand） or light conditions （in shade and open condition）. The results show that application of IAA and IBA induced more numbers of cuttings （collected from 10-year-old mother trees） to root compared to control. Branch cuttings of D. sissoo collected from 10-year-old mother trees and planted in soil bed in open conditions had 100.0% of cuttings to root in IAA （100 mg·L^-1） and IBA （200 mg·L^-1） treatments. Both rooting medium （Soil and sand） influenced significantly （p〈0.05） on rooting response of branch cuttings. Soil medium was found to achieve maximum no. of branch cuttings to root, compared to sand medium.

NaCl胁迫对降香黄檀(Dalbergia oderifera)种子萌发及幼苗生长的影响

设计5个不同NaCl浓度梯度(清水ck、1‰、2‰、3‰、4‰)试验,研究不同NaCl浓度梯度下降香黄檀种子萌发以及幼苗生长的差异。结果发现,随着NaCl胁迫程度的增强,降香黄檀种子萌发率逐渐下降、萌发时间推迟,幼苗的生长受到的抑制作用越强。

Polysaccharide,proline,and anti-oxidation enzyme activities of Thailand rosewood(Dalbergia cochinchinensis)seedlings exposed to exponential fertilization

Thailand rosewood(Dalbergia cochinchinensis)is a vulnerable high value tree species in subtropical and tropical Southeast Asia.The demand for the rehabilitation of the natural habitat of the species requires high-quality seedlings.However,current fertilizer regimes tend to deliver a coarse rate of fertilizers that result in a low uptake efficiency and a high risk of nutrient excess.Exponential fertilization is an effective approach to promote seedling quality through identifying an optimum dose out of a range of applications.In this study,Thailand rosewood seedlings were cultured in a mixed substrate of perlite,vermiculite,and peat(2:2:3,v/v/v)and received exponential fertilization from 0(control),through 100,200,300,400,and 600 mg nitrogen(N)per plant over a four-month cultural rotation.Height,diameter,and shoot biomass increased with fertilizer rate and were maximized at 300 mg N per plant.This application also reduced nutrient deficiencies and avoided excess supply compared to higher rates.Polysaccharide,proline,and anti-oxidation enzyme activities tended to be greater in unfertilized controls or in over-fertilized high dosages,with lower activities around 100–300 mg N plant(-1).Application of 300 mg N per plant was identified as the optimum dosage as it maximized nutrient reserves,polysaccharide and proline contents,and anti-oxidants without causing any symptoms from excess fertilizer toxicity.Future work is required to identify the effects of N-fixation on the formation of nutritional reserves in Thailand rosewood seedlings subjected to exponential fertilization.

Assessment of genetic diversity in Dalbergia sissoo clones through RAPD profiling

We studied the genetic polymorphism among 29 clones of shisham （Dalbergia sissoo Roxb） belonging to different geographic regions using random amplified polymorphic DNA （RAPD） markers. Out of 30 primers used, only 20 primers generated polymorphism in amplified product. In total 232 bands were amplified with 20 primers, of which 192 （82%） were polymorphic with an average of 9.6 bands/primer. The resolving power （Rp） ranged from 2.14 （Primer 5） to 11.93 （Primer 4）. Primer 4 and Primer 3 possessed high Rp value. Polymorphism information content （PIC） ranged from 0.15 （Primer 5） to 0.37 （Primer 4）. Primer 4 amplified total 18 bands in 29 genotypes with PIC value of 0.37 hence; this set of primer was most informative. The similarity coefficient analysis revealed two clusters. The first cluster comprised of only 10 clones and the second major cluster comprised of 19 clones. The genetic similarity among 29 clones ranged from 25.86% （clone 10 and 235） to 100% （clone 19 and 59）, suggesting a wide genetic base in shisham clones.

Effects of phosphorus and potassium addition on growth and nodulation of Dalbergia sissoo in the nursery

A study was conducted to test the influence of different inorganic fertilizers （phosphorous and potassium） on the nodulation and growth of Dalbergia sissoo grown in the nursery. Before seeds sowing, different combinations of P, K fertilizers were incorporated with the nutrient-deficient natural forest soils, and then amended with cowdung （soil： cowdung = 3：1）. Nodulation status （nodule number, shape, fresh weight, dry weight and color） in the roots and the plant growth parameters （length of shoot and root, collar diameter, fresh and dry weight of shoot and root） were recorded 60 days after seeds sowing. Nodulation status and growth of the plants varied significantly （P〈0.05） in the soils amended with fertilizers in comparison to the control. The highest nodule number （62）, fresh （0.50 g） and dry （0.07 g） weights were recorded with the dose of PK at the rate of 160 kg/hm2. Nodule shape and color also varied widely in different treatments. In case of plant growth parameters, shoot and root length, collar diameter, fresh and dry weight of the plants took on a significant difference （P〈0.05） among various combination of fertilizers. From the study, it is revealed that PK at the rate of 160 kg/hm^2 fertilizer with soil and cowdung mixture （soil： eowdung = 3：1） is recommended for optimum growth and nodule formation of D. sissoo in degraded soils at a nursery level.

Investigation of antioxidant and anti-inflammatory activity of leaves of Dalbergia paniculata(Roxb)

Objective:To investigate the antioxidant and anti-inflammatory activity of orally administered methanolic leaf extract of Dalbergia paniculala(D.paniculata) in Carrageenan induced inflammation in rats.Methods:In vitro antioxidant activity was evaluated for superoxide radical, Hydroxyl radical and DPPH radical scavenging activity.Three doses 200 mg/kg,400 mg/kg and 800 mg/kg of D.paniculala were tested for anti-inflammatory activity in Carrageenan induced rat paw edema model and paw thickness was measured every one hour up to 6 h. Results:The melhanolic leaf extract of D.paniculala produced dose dependent inhibition of Superoxide radical,Hydroxyl radical and OPPH radicals.In Carrageenan induced inflammation model,all three doses produced significant percentage inhibition of rat paw edema and 800 mg/kg dose produced maximum percent inhibition of rat paw edema(47.83%) at 3h compared to control group.Conclusions:In the present study we found that methanolic leaf extract of D.paniculata showed good in vitro antioxidant activity and in vivo anti-inflammatory activity in rats.

Variability in seed traits and genetic divergence in a clonal seed orchard of Dalbergia sissoo Roxb.

The variations in seed and pod traits, genetic superiority and genetic divergence were evaluated for a Clonal Seed Orchard （CSO） of Dalbergia sissoo Roxb. at Bithmera, India consisting of 20 clones from different agro-climatic conditions of four northern states （Uttar Pradesh, Uttarakhand, Haryana and Rajasthan）. The seeds and pods of various clones in the orchard exhibited significant variability in size, weight and other characters. Significant positive correlations were observed between seed length and seed width （p〈0.05）, seed length and sccd thickness （p〈0.01）, seed length and seed weight （p〈0.01）, seed thickness and seed weight （p〈0.01）, seed length and germination value （p〈0.05）. The ge- netic parameters for seed and pod traits also showed a wide range of variations in the orchard. Heritability values were found to be over 50 percent for most of ~e seed and pod traits. Seed weight, seed length and seed thickness showed high heritability values coupled with maximum genetic gain for these characters. Ward＇s minimum variance dendrogram of clones of D. sissoo showed three distinct clusters; cluster I was the largest with 12 better clones whereas cluster 2 and 3 consisting of seven moderate clones and one poor clone, respectively. Mean cluster values showed sufficient variation among the clusters for seed weight, germina- tion value and seed length. The possible hybridization between best clones of cluster I to the disease resistant clone of cluster 2 （resistant against deadly Gandoderma lucidum root rot disease of D. sissoo） is also suggested for further breeding programmes of the species. The deploy- ment of clone 194 （better performed and disease resistant） is also rec- ommended in future plantation programmes of D. sissoo in northern India.

Nursery practice on seed germination and seedling growth of Dalbergia sissoo using beneficial microbial inoculants

Nursery practice using microbial inoculants was performed to find out the efficacy of the inoculants on seed germination and seedling growth of sissoo（Dalbergia sissoo Roxb.）.Microbial inoculants or effective microorganisms（EM） are a mixture of many different beneficial microorganisms in a solution.The seedlings were grown in a mixture of sandy soil and cowdung（3：1） kept in polybags with pouring EM solution at different concentrations（0.1%,0.5%,1%,2%,5% and 10%） before and after a week of sowing the seeds.Seed germination rate and growth parameters of seedlings were measured,such as,shoot and root length,vigor index,fresh and dry weight of shoot and root and total biomass increment.The nodulation status influenced by EM was also observed along with the measurement of pigment contents in leaves.The highest germination rate（69%） was observed in 2% EM treatment,followed by 67% and 65% in 1% and 5% EM.The highest shoot length（33.2 cm） was in 2% EM,whereas highest root length（26.3 cm） was in 1% EM.Both fresh and dry weights from shoot and root,were maximum（4.16 g and 1.57 g;2.12 g and 0.83 g respectively） in 2% EM and were significantly（p ≤ 0.05） different from control.Vigor index was highest（4071） in 2% EM,which was significantly（p ≤ 0.05） different from control.Total dry weight increment was highest in 2% EM treatment,followed by 1% and 5% concentrations of EM.Nodulation number was higher at very low（0.1%） concentration of EM solution but it normally decreased with the increase of EM concentrations.The contents of chlorophyll a,chlorophyll b and carotenoid were highest（60.11,17.05 and 42.48 mg·L1respectively） in 2% EM treatment and lowest（39.35,13.55 and 27.29 mg·L-1 respectively） in control treatment.Therefore,low concentration of EM（up to 2%） can be used for getting maximum seed germination rate and seedling development of Dalbergia sissoo Roxb.

Present Situation and Recommendations on the Development of Dalbergia odorifera in Hainan

Dalbergia odorifera is particularly scarce,and the D. odorifera furniture and commercial raw materials on the market are mainly old furniture and agricultural tools,as well as commercially collected D. odorifera logs. The logs and high-grade furniture made from D. odorifera are extremely expensive. It takes several decades for D. odorifera to grow into wood. At present,there is only a small area of newly planted and regenerated forest,and the planting area of D. odorifera is about 15 000 ha in Hainan Island,distributed in the low altitude area of Hainan Island. It is suggested that Hainan D. odorifera island should be built through the transformation of low-yielding forest land,the use of wasteland and marginal cultivated land,land around houses,green belt and roadside land for the cultivation of D. odorifera,and the development area can be up to 100 000-150 000 ha. It is necessary to mobilize farmers to revitalize the rural green economy,plant D. odorifera,beautify the environment,and increase the economic resources of rural trees; actively publicize the legal nature and obligation of D. odorifera protection,and create a strong atmosphere for each family to plant and protect D. odorifera.

Genetic divergence for growth and wood parameters in different clones of Dalbergia sissoo Roxb

The wood analysis for different parameters was carried out in a clonal seed orchard ofDalbergia sissoo Roxb. established during 1997 at Hoshiarpur, Punjab, India. Twelve clones with higher index value were subjected to Euclidean Cluster Analysis based on wood and growth parameters to group into seven clusters. Cluster I and II contained four and three clones, respectively, and remaining clusters bad just one clone each. Clone originated from Barielly, Uttar Pradesh of cluster VII was found to be the most divergent clone. Cluster II with three clones main- tained greater inter cluster distance with other clusters. The divergence analysis has confirmed that the clones planted in the clonal seed orchard are sufficiently divergent and seed harvested from the orchard would maintain high diversity.

D2 analysis for estimating genetic divergence in different clones of Dalbergia sissoo

Genetic divergence was studied in selected 36 genotypes of Dalbergia sissoo Roxb. on the basis of seven morphological parameters. The divergence among genotypes was estimated by Mahalanobis method and genotypes were grouped into clusters by Tocher＇s method. All the genotypes were classified into seven distinct clusters on the basis of seven morphological traits. Cluster 1 was the largest with 25 genotypes followed by Cluster 2 （four genotypes）. Cluster 3, 5, 6 and 7 were the divergent clusters. The D2 analysis revealed that D2 value （39.42） between clone 5040 and clone 201 was recorded maximum. The intra-cluster distance ranged from 0.00 （Cluster 3, 5, 6 and 7） to 3.89 （Cluster 1）, the Cluster 3 （clone 33） was the most divergent cluster with maximum inter cluster distance （13.97） with the Cluster 7. By the divergence analysis, the parents for hybridization from diverse clusters could be selected for heterotic hybrids.

Genotype × environmental interactions for analyzing adaptability and stability in different clones of Dalbergia sissoo Roxb.

In total 36 superior clones of Dalbergia sissoo Roxb., screened from 300 selections conducted in natural and growing range of India and Nepal, were multiplied using single nodal cuttings and estab- lished to evaluate genotypexenvironmental interactions for adaptability and stability at the age of 30 months in three geographical locations in the state of Punjab, India. Clone 124 had maximum adaptability and stability （bi = 1.04） to perform exceedingly well over the locations. Clones 36 and 1 were stable with mean regression coefficient of 0.84 and 1.22, respectively. Nonetheless, clone 4 1 performed exceedingly well for all the characters to attain maximum population mean, and the perform- ance varied substantially across the locations. Therefore, clone 41 was considered as productive but non-adaptive clone. Though some of the clones were sensitive to sites, 14 clones for height, 16 for collar diameter, 12 for DBH and 7 for volume were relatively un-sensitive with higher regression coefficient. Nonetheless, clone 124 was the most Stable with average bi value of 1.04 and productive, which could play an important role in future breeding and commercial deployment of stable and produc- tive planting stock of Dalbergia sissoo.

Growth and heritability estimates among clones of Dalbergia sissoo Roxb. in a clonal seed orchard

The study of genetic variability is a prerequisite for any tree improvement program. Screening of clones showing the effect of dominance in growth and tree form is essential to identify productive clones, matched to sites. Our study performed in a clonal seed orchard at Lacchiwala of India, represented by 31 clones of Dalbergia sissoo （Shisham） Roxb. from diverse locations revealed significant variability at the 0.1% level of significance in various morphological traits. At the age of 13 years, clone 196 （Gonda） revealed maximum height of 20.2 m and a girth at breast height of 74.2 cm. The five identified clones viz. 196, 198, 192, 123 and 235 were well adapted to the climatic conditions of this zone, hence survived well and showed good performance compared to other clones. Heritability estimates of 87%, 83% and 80% for height, crown width and crown length respectively coupled with high genetic gains indicated strong genetic control of these variables. A correlation matrix of various growth attributes revealed positive correlation among these attributes. On the basis of the entire set of growth variables, the clones were grouped into two major clusters with three subgroups.

Vegetative propagation of Dalbergia sissoo:effect of growth regulators,length,position of shoot and type of cuttings on rooting potential in stem cuttings

The effect of auxins （IAA or IBA at 100, 200, 500 mg.L J） on rooting and sprouting parameters differed significantly （p 〈 0.05）. Rooting and sprouting percentages were higher in some treatments while other parameters were significantly higher in other treatments. The results of the present study revealed that rooting was significantly （p 〈 0.05） affected by the length of cuttings, as well as the position of shoots and type of cuttings. On average, 86.0 per cent rooting was observed in the 30-cm long cuttings com- pared to the other two lengths of cuttings, i.e., 20 and 25 cm, irrespective of any auxin treatment. In a control set, without any auxin treatment, a maximum of 60.0% rooting was recorded in the 30-cm long cuttings and a minimum of 40.0% of the cuttings rooted in the 20-cm long cuttings. Similarly, cuttings collected from the lower and upper portions of shoots resulted in 72.5 and 52.5 per cent rooting respectively, irrespective of auxin treatments. Maximum rooting was observed in the lower portion （40.0%） of cutting com- pared to the upper portion （30.0%） in the control set without auxin treatment. Leafless branches produced maximum （72.0%） rooting compared to the leafy cuttings （35.0%）, irrespective of the auxin treatment. The leafy cuttings produces a minimum of 15.0% sprout- ing and rooting in the control set.

Isolation of Two Furanoditerpenes and Two Triterpenes from the Stem Bark of <i>Dalbergia lanceolaria</i>L.f.

Stem bark of Dalbergia lanceolaria was subjected to methanolic extraction and extensive phytochemical separation. The process yielded four known compounds namely Jateorin (1), Columbin (2), β-sitosterol (3), and Lupeol (4). The compounds were identified using nuclear magnetic resonance spectral data namely 1H NMR, 13C NMR, 1H-1H COSY, HMBC, and DEPT-135 and verified by comparing the data with previous literature. All four compounds are being reported for the first time from the species D. lanceolaria. Furthermore, two furanoditerpenes namely Jateorin and Columbin are reported for the first time from the genus Dalbergia. This may lead to potential identification of a new chemotaxonomic marker for this genus.