Tissue Culture of Lespedeza cyrtobotrya

[Objective] The aim was to carry out study on tissue culture of Lespedeza cyrtobotrya. [Method] The seeds of L. cyrtobotrya were used as materials to study on its tissue culture. [Result] The best sterilization time to L. cyrtobotrya seeds was 8 min with 2.1% NaClO,in which shooting percent reached 37.8% and no polluted situations occurred. In the primary culture with the MS as basal medium,the concentration of 6-BA showed a significant effect on the index of buds differentiation,the optimum differentiation culture medium was MS＋BA 1.0 mg/L＋NAA 0.1 mg/L＋2,4-D 0.01 mg/L,on which the index of generation could reach 6.69. The optimum subculture medium was MS＋6-BA 1.0 mg/L＋2,4-D 0.05 mg/L. The plants can generate the highest roots and rooting percent with IBA 0.50 mg/L. [Conclusion] This study had provided theoretical basis for genetic improvement of L. cyrtobotrya.

木本植物适应酸性土壤机理的研究进展——以胡枝子(Lespedeza bicolor)和油茶(Camellia oleifera)为例

综述了近年来木本植物适应酸性土壤的耐铝机理研究进展,重点以酸性土壤先锋植物胡枝子(Lespedeza bicolor)和铝累积植物油茶(Camellia oleifera)为例,总结了木本植物根系有机酸的分泌、铝吸收和运输机制及铝与氮磷胁迫的协同适应等。木本植物有铝累积和铝排斥植物之分;铝排斥植物胡枝子耐铝的重要机制是其根系同时分泌柠檬酸和苹果酸;铝累积植物油茶高效累积铝的原因在于其不仅可以高效吸收土壤和土壤溶液中广泛存在的铝(Al3+和Al-F),而且可以通过木质部运输和特定季节韧皮部运输的配合实现铝的高效分配和传输;铵态氮相对于硝态氮可缓解胡枝子的铝毒害;磷对不同胡枝子耐铝作用的影响明显不同。木本植物适应酸性土壤机理的深入研究将会有助于完善植物的耐铝机理及铝运输理论,并为酸性土壤中矿质养分管理提供理论基础。

Structure, composition and enzymatic hydrolysis of steam-exploded lespedeza stalks

Pretreatment of lespedeza stalks by steam explosion has been studied. The results indicate that steam-exploded pretreatment has strong effects on physical features, morphology, crystallinity, and composition of lespedeza stalks as shown by scanning electron microscopy （SEM）, infrared （IR）, and X-ray diffraction spectrometry methods. After steam explosion, the cellulose and lignin contents of lespedeza stalks varied only slightly, but the hemicellulose content had decreased from 29.34% to 7.48%. The cellulose obtained by steam-exploded pretreatment had a higher degree of crystallinity than that of the raw material. At the explosion condition of 2.25 MPa and 4 min, lignocellulose is easier to hydrolyze by enzyme than the original lignocellulose. The concentration of reduced sugar in the hydrolyzate liquid increased from 71.77 to 162.84 g·L^-1.

水土资源恢复中的先锋豆科灌木──胡枝子（Lespedeza bicolor）的栽植研究

胡枝子栽植在侵蚀劣地上，其栽植技术简单，表现耐瘠、耐旱、耐寒，根系发达，生物量高，其枝叶或可作“三料”或可用以编织；同其它常见灌木相比，具有显著的生长优势。利用胡枝子碎粉培殖香茹，既能节约大量木材，又可获取可观经济效益；胡枝子栽植对侵蚀劣地蓄水改土效果明显，对恢复土壤肥力和水利资源，因而值得推广。

Simultaneous saccharification and fermentation of steam-pretreated lespedeza stalks for the production of ethanol

Lespedeza stalks were subjected to steam pretreatment at 210℃ for some steaming time before simultaneous saccharification and fermentation （SSF）. Cellulose-derived glucose was extensively utilized by yeast during SSE The ethanol yields after steam pretreatment of the lespedeza stalks at 210℃ were 59.3%, 72.8% and 62.2% of the theoretically expected values when the steaming times were 2, 4 and 6 min, respectively. The highest yield from α-cellulose was 92.7% of the theoretical value. Steam explosion pretreatment of lespedeza stalks increased ethanol yields by a factor of 4.4, from 16.4% （untreated） to 72.8% （steam explosion pretreated）.

Phenotypic variation in thirteen native provenances of Lespedeza bicolor Turcz

Lespedeza bicolor Turcz is a native shrub in the temperate region of China. Thirteen provenances of L. bicolor were investigated in our study. Morphological diversity among and within provenances were analyzed based on 19 phenotypic traits, including seven vegetative growth characteristics, nine floral traits and three pod characteristics. The coefficient of variation （CV） ranged from 6.4% to 65.4% and the phenotypic differentiation coefficient from 43.3% to 97.3%. Analyses of variance showed that there were extremely significant variations among and within provenances in pod length. Differences in plant height, inter-nodal length, number of branches, leaf width, leaf length and leaf area were not remarkable among provenance, but distinct within provenances. Other traits showed evidence of contrasts. Variation among provenances was the main part of phenotypic variation. Correlation analysis indicated that there were highly significant positive correlations between plant height and other vegetative growth characteristics. Floral traits were not associated with vegetative growth characteristics. The 13 provenances of L. bicolor investigated can be classified into three groups according to a UPGMA cluster analysis.

Effects of Four Chemicals on Tissue Differentiation of Lespedeza bicolor in In-vitro Culture

In order to find the optimal bacteriostatic agents and selectors and their concentrations in genetic transformation of Lespedeza bicolor with BADH as a marker gene, an resistance experiment was carried out with four chemicals, NaCl, betaine aldehyde, PEG and cefotaxime （Cef） added into the media, Different concentrations and combinations of different chemicals were added into the media in the stage of adventitious bud regeneration of cotyledonary nodes, Shoot segment multiplication, and rooting. The results showed that NaCI and betaine aldehyde could inhibit tissue differentiation of L. bicolor, and could thus be used as selectors. Furthermore, 0.8 g/L NaCI could completely inhibit the cotytedonary node differentiation and shoot segment multiplication, and 0.5 g/L NaCI was optimal for inhibition of rooting. It was also found that with the concentration increase of betaine aldehyde in media, adventitious buds regenerated from cotyledonary nodes decreased, and 1.5 g/L was confirmed to be the optimum concentration. Moreover, the mixture of 0.7 g/L NaCI and 0.3 g/L betaine aldehyde significantly inhibited shoot segment multiplication and rooting, so this mixture could be used asa selector. The Cef ex- periment showed that 200-300 mg/L C, ef in media did not inhibit significantly the regeneration of adventitious buds from cotyledonary nodes, and 100 mg/L Cef had little effect on the shoot segment multiplication and rooting.

Effect of Ensiling on Efficacy of Sericea Lespedeza against Gastrointestinal Nematodes and Coccidia in Goats

Effect of ensiled sericea lespedeza(SL,Lespedeza cuneata)on indicators of gastrointestinal parasitism was investigated in two trials using intact male Spanish goats(n=36/trial).Naturally parasite-infected animals at 24.4±3.7 kg body weight(BW)and nine months of age or 24.6±0.57 kg BW at 4-6 months of age were used for 28 d or 21 d for Trials 1 and 2,respectively.For Trial 1,goats were fed SL silage(SLS),SL hay(SLH),or Bermuda grass(BG,Cynodon dactylon)hay at 70%of the diet.For Trial 2,goats were provided with SLH or SLS and orally drenched with distilled water(Hay or silage plus water,HW and SW,respectively)or polyethylene glycol(PEG;SLS only;SP)daily.Fecal and blood samples were collected weekly to determine gastrointestinal nematode fecal egg counts(FEC)and coccidia fecal oocyst counts(FOC)per gram of feces and blood packed cell volume(PCV).The SL diets significantly reduced(p<0.05)FEC and FOC in both trials,with the SLH diet reducing FEC 7 d faster compared to SLS.In PEG-treated goats,FOC tended to increase(p<0.07)at day 7 before decreasing.There was no treatment effect on PCV scores.Overall,SLH and SLS reduced fecal egg and oocyst counts in goats,but PEG results were inconclusive,so more research is needed.

Effect of Ground and Pelleted Sericea Lespedeza Whole Plant and Leaf Only on Gastrointestinal Nematode and Coccidial Infection in Goats

Two 42-day feeding trials in pens were completed with young goats(Spanish,intact male,six months old,n=29 Trial 1;n=20 Trial 2)to determine the anti-parasitic bioactivity of whole plant and leaf only sericea lespedeza(SL;Lespedeza cuneata)fed as a ground meal or in pelleted form.In Trial 1,goats were fed SL leaf only pellets,SL whole plant pellets,or a commercial goat pellet as 50%of a complete ration,while in Trial 2,kids were fed either ground SL leaf meal or ground whole plant SL meal as 25%of a complete ration.Fecal samples were collected weekly for determination of gastrointestinal nematode(GIN)fecal egg counts(FEC)and coccidia fecal oocyst counts(FOC).In Trial 1,both of the pelleted SL rations reduced FEC(p<0.06)relative to the kids fed the control ration,and for FOC,there were lower values on Days 7 to 42 relative to Day 0 for goats on either pelleted SL diet,while FOC for control animals did not change over time(interaction,p<0.01).In Trial 2,both ground whole plant and leaf only SL diets reduced(p<0.05)FEC of the goats over time,but the effect was faster in the kids on the SL leaf meal ration.At 50%of the diet,SL leaf and whole plant meal pellets were equally effective against GIN egg and coccidial oocyst production in young kids,but SL leaf meal was more effective in reducing GIN egg production than whole plant SL meal when fed at 25%of the diet.

Effect of Supplemental Sericea Lespedeza Pellets on Internal Parasite Infection and Nutritional Status of Grazing Goats

Feeding pelleted sericea lespedeza （SL; Lespedeza cuneata） on pasture can reduce gastrointestinal nematode （GIN） and coccidia （Eimeria spp.） infection in sheep and goats, but effects on nutritional status are unclear. Therefore, a study was completed comparing yearling goats grazing grass pasture supplemented with SL or non-condensed tannins （CT） commercial pellets （control group）, respectively, at 1.5% of body weight for 14 weeks, and then after 14 weeks, they were only fed with non-CT pellets at 2.5% of body weight for additional six weeks. Animal body weight was measured at the start of the trial, the 7th week, 14th week and end of the trial. Fecal samples were taken weekly to determine GIN egg output （fecal egg count; FEC） and coccidial oocyst production （fecal oocyst count; FOC）. Blood samples （to determine packed cell volume; PCV） were taken weekly to monitor anemia status of the goats, and on days 0, 98 and 137 to determine aspartate aminotransferase （AST）, alanine aminotransferase （ALT） and creatine kinase （CK）. The study revealed that supplemental SL leaf meal pellets had no effect on FEC, but reduced FOC （P 〈 0.05） and improved FAMACHA~ scores （P 〈 0.001） in the goats, and the SL supplemented goats tended to gain more weight （P = 0.07） than control animals during the first 49 d, following initiation of pellet feeding. There were no treatment effects on enzymes related to liver function or muscle turnover （AST, ALT and CK）, suggesting that there was no muscle damage due to long-term feeding of SL pellets. In conclusion, feeding supplemental SL pellets at 1.5% of body weight on pasture may be a viable strategy for improving health and productivity of yearling goats.

Smart Man＇s Sericea Lespedeza and Worm Control： A Review

Sericea lespedeza （Lespedeza cuneata （Dum. Cours.） G. Don.） has historically been called ＂poor man＇s alfalfa＂, because of its ability to grow on marginal soil with the minimal inputs of lime or fertilizer. With recent research detailing the potential health benefits of this forage to animals, it may be time for a new nickname. Although sericea lespedeza has been used for soil conservation and as an inexpensive （low-input） source of pasture or hay crop for nearly 100 years, research over the last 10-15 years has demonstrated the excellent bioactivity of this plant against infection with gastrointestinal nematodes （Haemonchus contortus） and protozoan parasites （Eimeria spp.） in livestock. This bioactivity, which has been attributed to a unique type of condensed tannins （CT）, has been confirmed in both fresh （grazed） and dried （hay, meal, pellets） forms of sericea lespedeza in a number of studies with sheep, goats, and cattle. There is on-going research with this forage to determine the effect of ensiling on its bioactive properties and to validate its anti-parasitic effects on-farm. Future research will focus on determining the mode of action of sericea lespedeza tannins against internal parasites and the interplay between the nutritional value and the health benefits of this plant.

Effect of Ensiling on Nutritional Properties of Sericea Lespedeza Alone or in Mixtures with Alfalfa

In this investigation,fresh sericea lespedeza(SL;Lespedeza cuneata)and alfalfa(Medicago sativa)were cut in the field,frozen,chopped and mixed into ratios of 100:0,75:25,50:50,25:75 and 0:100,respectively,with each treatment combination packed into 12 mini-silos and sealed to be air-tight.Three mini-silos per treatment were opened after 1,7,21 and 84 d of ensiling and analyzed for pH,neutral detergent fiber(NDF),acid detergent fiber(ADF),and acid detergent lignin(ADL),unbound and bound condensed tannins(CT),nitrogen(N),nitrate N(NO3-N),and ammonia N(NH4+N)content.All of the forage combinations ensiled well,with a rapid drop in pH(below 5.0 by Day 7).Fiber concentrations(NDF,ADF,ADL)were greater in 75% and 100% SL silages than in 0%,25% and 50% SL samples by Day 84 of the study,possibly due to interference of CT in the detergent analysis system.Concentrations of N,NO3-N and NH4+N were decreased in silages as percentage SL in the mixture increased,while unbound,bound and total CT increased as percentage SL increased.In this study,there was reduced proteolysis during ensiling of combinations of SL and alfalfa,as indicated by reduced NO3-N and NH4+N production as percentage SL in the silage mixtures increased.

Short-term Intensive Sustainable Restoration of Grasslands and Prairies Invaded with High Densities of Nitrogen-fixing Weeds： A Test with the Invasive Plant Lespedeza Cuneata

This study examines a management strategy for restoring grassland and prairie communities that have become degraded due to high density stands of invasive nitrogen-fixing plants. The novel management applications minimize the use of herbicides and maximize the competitive interactions of native species. The management method includes two seasons of application of organic fertilizer （4-1-4）, an initial herbicide （Pasture Gard, Dow Agro） application, and mowing, where mowing was a necessary treatment to control secondary growth in prairie habitats, to control high density patches of Lespedeza （L.） Cuneata, in a completely randomized factorial experiment. The herbicide was effective in reducing L. Cuneate stem density 0 stems/m^2 from an initial 88 stems/m^2 with cover reduced to 0% from 16%. The fertilizer only treatment reduced L. Cuneata percent cover to 6% from initial cover of 16%, but did not reduce the number of stems. The management strategy is an effective fast step in restoring a native prairie invaded by a nitrogen-fixing plant.

达乌里胡枝子(Lespedeza davurica)混播下叶绿素荧光特性对水分变化的响应

为研究黄土丘陵区白羊草植物群落优势种叶绿素荧光动态变化特性对土壤水分变化的响应,采用盆栽控制试验,设置7个组合比例(白羊草∶达乌里胡枝子, 0∶12, 2∶10, 4∶8, 6∶6, 8∶4, 10∶2和12∶0)和3种水分水平(高水(HW),中水(MW)和低水(LW),对应为田间持水量的80%±5%, 60%±5%和40%±5%),并分别在达乌里胡枝子现蕾期、开花期与结实期将土壤水分从高水降到中水(H-MW)和低水(H-LW),测定了达乌里胡枝子在水分降低后第0天、第1天、第3天、第7天的叶绿素荧光参数。结果表明:在土壤水分阶段干旱胁迫后,最大光化学效率(Fv/Fm)和实际光化学效率(ФPSⅡ)总体上在开花期显著最高、结实期显著最低,且Fv/Fm变化范围为0.767~0.829,表明达乌里胡枝子具有更为稳定的光化学效率;现蕾期和开花期的ФPSⅡ均以H-MW显著高于H-LW,表明土壤水分降低幅度较大显著削弱PSⅡ反应中心的实际光能捕获效率;开花期的非光化学淬灭系数(NPQ)为H-LW显著高于H-MW,表明达乌里胡枝子开花期通过耗散过剩激发能以减轻重度水分胁迫对叶片光合结构的损伤;单播下的Fv/Fm、ФPSⅡ和光化学淬灭系数(q P)显著高于或与混播无显著差异,表明混播对土壤水分阶段降低后达乌里胡枝子的光合反应过程产生抑制作用。本研究为阐明土壤水分变化条件下两草种竞争与共生的生理生态机制提供理论依据。

Methanolic extract from Lespedeza bicolor： potential candidates for natural antioxidant and anticancer agent

OBJECTIVE: To evaluate the anticancer, antioxidant and antimicrobial activities along with total phenolic and flavonoids contents extracted from Lespedeza bicolor indigenous to Khyber Pukhtoonkhwa Pakistan.METHODS: Anticancer activity against human lung carcinoma(LU-1) and human prostate carcinoma(Ln Cap), antimicrobial against Aspergillus fumigates, Aspergillus niger, Fusarium solani, Mucor sp Klebsiella pneumonia and Enterococcus and antioxidant activity on 1,1-Diphenyl-2-picryl-hydrazy(DPPH) and Hydrogen peroxide radicals scavenging activity were assessed in methanol extract o Lespedeza bicolor.RESULTS: Lespedeza bicolor aerial parts and root extract showed IC_(50)value of 12.5 and 50 μg/m L against LU-1 whereas, ≤ 12.5 and 12 μg/m L were calculated against Ln Cap cell line. Minimum inhibition concentration(MIC) value of 20-35 μg/m L has been observed against Aspergillus fumigates, Aspergillus niger, Fusarium solani and Mucor sp in comparision with 1-2.5 μg/m L of Terbinafine used as a standard fungicide. MIC value of 20 and 35 μg/m L of Lespedeza bicolor aerial parts and root extract against bacterial pathogen Klebsiella pneumonia and 20-50 μg/m L against Enterococcus has been measured. DPPH radical scavenging activity of Lespedeza bicolor with IC_(50)values of ≤ 50 μg/m L and ≤ 200 μg/m L was observed whereas, hydrogen peroxide scavenging activity with IC_(50)values of ≤25 μg/m L for aerial parts and ≤ 50 μg/m L for the root extract of Lespedeza bicolor has been shown.CONCLUSION: These data suggested that the methanolic extract of Lespedeza bicolor could be potential candidates for natural antioxidants and anticancer.

Study of Ultra-drying on Vigor and Physiological Characteristics of Lespedeza bicolor Seeds

Ultra-drying (in a desiccator with silica gel) and accelerated aging (50 oC, 30 d) of Lespedeza bicolor seeds from the arid and semiarid areas of northwest China were studied. The results showed that in the moisture content of seed ( 43?70 g/kg), germination percentage and vigor indices had no significant differences; ultra-dried seeds had a stronger anti-aging ability than those with natural drying, and the best MC for the storage was 43 g/kg. Therefore, it was feasible to store L. bicolor seeds with ultra-dried moisture content.

Two new phenylpropanoid glycosides from the aerial parts of Lespedeza cuneata

Two new phenylpropanoid glycosides named cuneataside E (1) and cuneataside F (2), were isolated from the aerial parts of Lespedeza cuneata (Dum. Cours.) G. Don, whose structures were E and Z isomer, respectively. Their structures were elucidated on the basis of comprehensive spectroscopic analysis (UV, IR, HR-ESI-MS, 1D and 2D NMR). In in vitro bioassays at 10 mu mol/L, compound 1 showed moderate hepatoprotective activity against N-acetyl-p-aminophenol (APAP)-induced toxicity in HeG2 cells. (C) 2016 Chinese Pharmaceutical Association and Institute of Materia Medica, Chinese Academy of Medical Sciences. Production and hosting by Elsevier B.V. This is an open access article under the CC BY-NC-ND

达乌里胡枝子根瘤菌的促生作用及高效根瘤菌的鉴定

本研究选择达乌里胡枝子(Lespedeza daurica)根瘤分离且结瘤效果较好的7株根瘤菌(TG64,TG97,TG98,TG100,TG101,TG105和TG115),通过测定达乌里胡枝子生长表型和主要营养成分,结合结瘤和光合特性初步分析促生的生理生化机制,研究不同根瘤菌的促生作用;通过分子生物学、形态学和生理生化特性鉴定高效根瘤菌。结果表明:除TG64,其余菌株显著提高了株高、根长、生物量和蛋白质含量,改善了根系构建,TG101促生最明显。所有处理单株结瘤5~33个,根瘤干重2.55~35.87 mg;TG98,TG100,TG101和TG115显著提高了叶绿素含量、光合和氮吸收能力,是促生的主要生理生化机制。经鉴定,TG101为格木慢生根瘤菌(Bradyrhizobium erythrophlei)。综上,根瘤菌通过改善植株氮营养促进叶绿素合成、增强光系统活性,提高光合能力,进而促进干物质积累调控生长。其中TG101为高效根瘤菌,为后续研发适用黄土高原的达乌里胡枝子根瘤菌剂提供了菌种资源和理论依据。

硫化氢对盐胁迫下达乌里胡枝子生长和抗氧化系统的影响

为探讨外源硫化氢对盐胁迫下达乌里胡枝子(Lespedeza davurica)幼苗生长和抗氧化系统的影响,本试验以盐胁迫下的达乌里胡枝子为研究对象,通过施加不同浓度的外源硫化氢,并在盐胁迫第0,10,20,30 d分别测定幼苗的生长特性和抗氧化系统指标。结果表明:施加外源硫化氢缓减了盐胁迫对达乌里胡枝子幼苗生长的抑制作用,提高了盐胁迫下达乌里胡枝子幼苗抗氧化酶活性以及抗坏血酸和谷胱甘肽含量,增加了叶片相对含水量,降低了叶片相对电导率以及丙二醛的含量。综上所述,施加外源硫化氢在一定程度上提高了达乌里胡枝子幼苗叶片的抗氧化酶活性,延缓了膜脂过氧化的发生,因此增强了达乌里胡枝子幼苗对盐的耐受性。

达乌里胡枝子愈伤组织及种子EMS诱变研究

为优化达乌里胡枝子愈伤诱导培养基的激素含量及比例,明确甲基磺酸乙酯(EMS)诱变愈伤组织及种子的最佳条件,本研究以达乌里胡枝子种子为外植体,探究了不同激素配比对愈伤诱导的影响,并针对愈伤组织和种子处理分别设置了两时间两浓度(2 h和6 h,0.3%和0.9%)及三时间两浓度(6 h、8 h、10 h,0.4%、0.8%)的EMS处理时间和浓度的组合,研究各处理对愈伤分化系数和后期植株成活率以及种子发芽率、发芽势和幼苗表型变异的影响。结果表明,达乌里胡枝子种子愈伤的最佳激素配比为0.25 mg/L NAA+0.5 mg/L 6-BA+0.5 mg/L 2,4-D,此时达乌里胡枝子愈伤诱导率为94.9%。EMS诱变达乌里胡枝子愈伤组织的适宜浓度为0.3%,处理时间为6 h,分化系数和植株相对成活率分别为46.31%和47.78%,接近半致死范围,且生根数、根长和平均株高等指标与高浓度EMS处理相比受到的负面影响较小。EMS诱变达乌里胡枝子种子的适宜浓度为0.8%,处理时间10 h,发芽率、出苗率、成苗率分别为53.61%、51.06%和53.28%,均接近半致死量,同时发现多株幼苗叶片出现了明显的形态变异。本试验在达乌里胡枝子原有种质资源基础上,丰富了达乌里胡枝子突变体库,也为后续开展达乌里胡枝子诱变育种研究提供理论依据。