Cloning and sequence analysis of a glucose-6-phosphate dehydrogenase gene <Emphasis Type="Italic">PsG6PDH</Emphasis> from freezing-tolerant <Emphasis Type="Italic">Populus suaveolens</Emphasis>

A 1207 bp cDNA fragment (PsG6PDH) was amplified by RT-PCR from cold-induced total RNA of the freezing-tolerant P. Suaveolens, using primers based on the highly conserved region of published plant glucose-6-phosphate dehydrogenase (G6PDH) genes. The sequence analysis showed that PsG6PDH coding region had 1 101 bp and encoded 367 predicted amino acid residues. Moreover, the nucleotide sequence of PsG6PDH showed 83%, 82%, 79%, 79% and 78% identity, and the derived amino acid sequence shared 44.2%, 44.7%, 42.0%, 40.5% and 43.9% identity with those of the Solanum tuberosum, Nicotiana tabacum, Triticum aestivum, Oryza sativa and Arabidopsis thaliana, respectively. The results show that PsG6PDH is a new member of G6PDH gene family and belongs to the cytosolic G6PDH gene. This is the first report on cloning of the G6PDH gene from woody plants.     

High-level expression of 4-coumarate:coenzyme A ligase gene Pt4CL1 of Populus tomentosa in E. coli

In order to investigate the enzymatic properties of the 4CL1 of Populus tomentosa, the recombinant expression vector pQE31-4CL1 was constructed. The recombinant was identified by three restriction endonucleases, then the vector pQE31-4CL1 was transformed into expression host M15 (pREP4) and induced by isopropyl-α-D-thiogalactoside (IPTG) to express 60 kD fused protein Pt4CL1. The biologically active Pt4CL1, expressed as soluble protein, was achieved with 0.6 mmol·L-1 IPTG induction as the ex-pression temperature declined from 37 to 28°C. The 6×His tag facilitates affinity binding to Ni2 -nitrolotriacetic acid (NTA) and enables one-step purification to acquire the molecular SDS-PAGE electrophoresis purity of the active 4CL1 protein by agarose cou-pled with Ni2 -NTA affinity chromatography. The optimal substrate for Pt4CL1 was 4-coumarate.     

At5g54160 gene encodes <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> 5-hydroxyconiferaldehyde <Emphasis Type="Italic">O</Emphasis>-methyltransferase

The function of an Arabidopsis thaliana gene, At5g54160 annotated as a caffeic acid O-methyltransferase CAOMT gene was characterized. The recombinant enzyme of this gene (AtOMT1) catalyzed the O-methylation of phenylpropanoid and flavonoid substrates. The specificity constants (k _cat/K _m) for 5-hydroxyconiferaldehyde (5-HCAld) and quercetin were both 0.11 μM⁻¹·min⁻¹. On the other hand, lignins of At5g54160-knockout Arabidopsis mutants lacked syringyl units. In addition, we showed that the gene silencing also resulted in significant accumulation of caffeyl alcohol (CaAlc). These results strongly suggested that At5g54160 gene is involved in syringyl lignin synthesis for the methylation of both 5-hydroxyconiferaldehyde and 3,4-dihydroxyphenyl compound(s). Part of this work was presented at the Annual Meeting of Japan Society for Bioscience, Biotechnology, and Agrochemistry, March 24–27, 2007     

Management Effects on Biomass and Foliar Nutritive Value of <Emphasis Type="Italic">Robinia pseudoacacia</Emphasis> and <Emphasis Type="Italic">Gleditsia triacanthos</Emphasis> f. <Emphasis Type="Italic">inermis</Emphasis> in Arkansas,USA

The browse potential of black locust (Robinia pseudoacacia L.) and thornless honey locust [Gleditsia triacanthos f. inermis (L.) Zabel] has not been adequately tested. Our objective was to determine effects of fertilization and pollarding on biomass and foliar nutritive value in separate studies of black locust and thornless honey locust in Arkansas, USA. Shoots were sampled monthly for two consecutive growing seasons in 2002 and 2003 to determine foliar, shoot, and total aboveground biomass, shoot basal diameter, and foliar nutritive value (crude protein and in vitro digestibility). Black locust yielded more foliar biomass when pollarded at 50 or 100 cm and fertilized with 600 kg P ha⁻¹, than at 5 cm with or without P, averaging 3.5 Mg dry matter ha⁻¹. Black locust foliar crude protein and in vitro dry matter digestibility ( ≤ 170 and 534 g kg⁻¹, respectively) decreased as leaves aged, but still met maintenance needs for beef cattle (Bos taurus L.). Thornless honey locust had little agronomic potential because of slow establishment, low foliar yield (330 kg ha⁻¹), and a 2% reversion to undesirable thorny phenotype. Black locust should be considered for livestock browse when drought induces semi-dormancy of herbaceous forages.     

Successful <Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated transformation of <Emphasis Type="Italic">Populus tomentosa</Emphasis> with apple <Emphasis Type="Italic">SPDS</Emphasis> gene

The problem of salinized soils has become one of the most serious constraints to agricultural and forest productivity. With the purpose of enhancing salt stress tolerance of Populus tomentosa, we transformed this tree species with spermidine synthase （SPDS） genes derived from an apple by an Agrobacterium-mediated method. Four transgenic clones were confu＇med by PCR and Southern blot analysis. As well, the expression of introduced SPDS genes was analyzed by real-time quantitative PCR.     

<Emphasis Type="Italic">In vitro</Emphasis> morphogenic response of leaf sheath of <Emphasis Type="Italic">Phyllostachys bambusoides</Emphasis>

Nodal segments from secondary branches of saplings of Phyllostachys bambusoides were inoculated in MS medium to assess the in vitro morphogenic response of leaf sheath through the induction to callogenesis by Picloram(4-amino-3,5,6-trichloropicolinic acid) at different concentrations of carbohydrate under the same conditions with presence or absence of luminosity.In our experiment,secondary explants were kept in MS medium containing 8.0 mg·L-1 of Picloram for the callus formation.Calluses were transferred in MS medium supplemented with sucrose,fructose and glucose(control,2%,4% and 6%).Results show that Picloram induced the callogenesis in leaf sheath.The secondary embryogenesis was formed in yellow-globular callus.The sucrose as carbohydrate source in the absence of light was more efficient to induce rhizogenesis.Glucose was more efficiency in the presence of light.Callogenic induction and further embryogenesis evidenced the competence and determination of leaf sheath cells.     

Gas exchange responses of <Emphasis Type="Italic">Eucalyptus</Emphasis>, <Emphasis Type="Italic">C. africana</Emphasis> and <Emphasis Type="Italic">G. robusta</Emphasis> to varying soil moisture content in semi-arid (Thika) Kenya

A dramatic decline in forest cover in eastern Africa along with a growing population means that timber and poles for building and fuelwood are in short supply. To overcome this shortage, the region is increasingly turning to eucalyptus. But eucalyptus raises environmental concerns of its own. Fears that it will deplete water supply, affect wildlife and reduce associated crop yields have caused many countries in the region to discourage farmers from planting this exotic. This paper is part of a series of investigations on the growth and water use efficiency of faster growing eucalyptus hybrids, which was introduced from South Africa to Kenya. The hypothesis is that the new hybrids are more efficient in using water and more suitable for the semi-arid tropics than existing eucalyptus and two popular agroforestry species. Gas exchange characteristics of juvenile Eucalyptus grandis (W. Hill ex Maiden), two eucalyptus hybrids (E. grandis × Eucalyptus camaldulensis Dehnh.), Grevillea robusta (A. Cunn) and Cordia africana (Lam) was studied under field and pot conditions using an infrared gas analyzer was used to measure photosynthetic active radiation (PAR), net photosynthetic rate (A), stomatal conductance (g _s) and transpiration rate (E) at CO₂ concentrations of 360 μmol mol⁻¹ and ambient humidity and temperature. A, E and g _s varied between species, being highest in eucalyptus hybrid GC 15 (24.6 μmol m⁻² s⁻¹) compared to eucalyptus hybrid GC 584 (21.0 μmol m⁻² s⁻¹), E. grandis (19.2 μmol m⁻² s⁻¹), C. africana (17.7 μmol m⁻² s⁻¹) and G. robusta (11.1 μmol m⁻² s⁻¹). C. africana exhibited high E values (7.0 mmol m⁻² s⁻¹) at optimal soil moisture contents than G. robusta (3.9 mmol m⁻² s⁻¹) and eucalyptus (5.3 mmol m⁻² s⁻¹) in field experiment and G. robusta (3.2 mmol m⁻² s⁻¹) and eucalyptus (4.2 mmol m⁻² s⁻¹) in pot-grown trees. At very low soil moisture content, extremely small g _s values were recorded in GC 15 and E. grandis (8 mmol m⁻² s⁻¹) and G. robusta (14 mmol m⁻² s⁻¹) compared to GC 584 (46.9 mmol m⁻² s⁻¹) and C. africana (90.0 mmol m⁻² s⁻¹) indicating strong stomatal control by the species. Instantaneous water use efficiency ranged between 3 and 5 μmol mmol⁻¹ and generally decreased with decline in soil moisture in pot-grown trees but increased with declining soil moisture in field-grown trees.     

Laboratory and field evaluation of <Emphasis Type="Italic">Beauveria bassiana</Emphasis> for controlling Mulberry whitefly <Emphasis Type="Italic">Pealius mori</Emphasis> Takahashi (Homoptera: Aleyrodidae) in mulberry (<Emphasis Type="Italic">Morus alba</Emphasis> Linn)

In a laboratory screening of 12 isolates of entomopathogenic fungi against nymphs of the mulberry whitefly (Pealius mori Takahashi), Beauveria bassiana (Balsamo-Crivelli) Vuillemin CKB-048 was the most virulent, causing 87 ± 3% mortality at 1 × 10⁶ conidia/ml. Infection was confirmed by growth of the fungus from cadavers and by scanning electron microscopy of treated nymphs. Beauveria bassiana CKB-048 was formulated as a wettable powder (1 × 10⁹ conidia/g) and tested in two mulberry (Morus alba Linn) plantations in central and northeastern Thailand. In both locations, two spray applications of B. bassiana CKB-048 at 3.75 × 10¹² to 6.25 × 10¹² conidia/ha and at 14 day intervals provided good control of whitefly nymphs; control with B. bassiana CKB-048 was comparable to that with the pesticide buprofezin at 250 g of active ingredient/ha. In addition, no mortality of silkworm larvae occurred when the larvae were fed with mulberry leaves sprayed with B. bassiana CKB-048 7, 14, or 21 days earlier.     

Insecticidal efficacy of <Emphasis Type="Italic">Isaria farinosa</Emphasis> in different life stages of <Emphasis Type="Italic">Pissodes punctatus</Emphasis> (Coleoptera: Curculionidae)

The insecticidal activity of an entomopathogenic fungus, Isaria farinosa on pine bark-weevil (Pissodes punctatus) at different life stages underwent laboratory testing. Larvae, pupae and adults of Pissodes punctatus were exposed to a range of concentrations of fungal conidia in suspension for 5 s; mean mortality and LC₅₀ values were calculated. The results showed I. farinosa can effectively infect larvae, pupae and adult pine bark-weevils, and the highest mean mortalities at each life stage can exceed 88%. The mean mortalities increased with higher concentrations of I. farinosa. Larvae were most susceptible with LC₅₀ 1.2 × 10⁶ conidia ml⁻¹ (15 days), pupae (30 days) exhibited the second largest effects, and adults (15 days) with LC₅₀ 1.72 × 10⁶ and 1.99 × 10⁶ conidia ml⁻¹, respectively, at end of the experiments. This suggests that I. farinosa could be a potential bio-control agent against the pine bark-weevil.     

Warfarin resistance test and polymorphism screening in the <Emphasis Type="Italic">VKORC1</Emphasis> gene in <Emphasis Type="Italic">Rattus flavipectus</Emphasis>

The buff-breasted rat (Rattus flavipectus) is a major agricultural pest across China. Warfarin-resistant animals have been found in several major provinces in China, and are hampering effective control. Molecular mechanisms underlying resistance to anticoagulant rodenticides have been determined for other species, but genetic information regarding resistance in R. flavipectus remains unknown. The vitamin K 2,3-epoxide reductase complex subunit 1 (VKORC1) encoded by VKORC1 gene is the molecular target of coumarin anticoagulants, and amino acid substitutions in VKORC1 coding-regions have been reported as one of the supposed mechanisms of warfarin resistance. Here, lethal feeding test in R. flavipectus (n = 36) was conducted in Zhanjiang, China. Four animals (11%) survived the test period of 25 days and were identified as warfarin resistance. Polymorphism across the whole genome DNA sequence of the VKORC1 gene was screened out and compared with resistant and non-resistant rats. A total of nine single nucleotide polymorphisms (SNPs) were identified including seven SNPs in introns and two SNPs in exons, and the SNP (2317A > G) located in exon 3 led to the amino acid substitution (Tyr139Cys) in VKORC1 protein. Based on the characteristics of Tyr139Cys mutation of VKORC1 in humans or rats and its relationship with warfarin-resistance, Tyr139Cys mutation may be one mutation responsible for anticoagulant resistance in R. flavipectus. Given the low numbers of resistant rats in our feeding test, wider surveillance, tests of resistance development in a larger wild population and further researches on the genetic mechanisms of anticoagulant resistance in R. flavipectus are necessary.     

Insecticidal properties of <Emphasis Type="Italic">Thymus persicus</Emphasis> essential oil against <Emphasis Type="Italic">Tribolium castaneum</Emphasis> and <Emphasis Type="Italic">Sitophilus oryzae</Emphasis>

Red flour beetle Tribolium castaneum (Herbst) and rice weevil Sitophilus oryzae (L.) are considered to be the major insect pests in storage. Essential oils from aromatic plants are recognized as proper alternatives to fumigants. Thymus persicus (Ronniger ex Rech. f.) is one of these plants that have medicinal properties and is indigenous to Iran. The essential oil was obtained from aerial parts of the plant and analyzed by GC and GC–MS. Carvacrol (44.69%) and thymol (11.05%) were the major constituents of the oil extracted. In this experiment, fumigant toxicity of the essential oil was studied against T. castaneum, S. oryzae at 27 ± 1°C and 60 ± 5% RH in dark condition. The adult insects were exposed to the concentrations of 51.9, 111.1, 207.4 and 370.4 μl/l air to estimate median lethal time (LT₅₀) values. The fumigant toxicity was increased in response to increased essential oil concentrations. The LT₅₀ values at the lowest and the highest concentrations tested were ranged from 28.09 to 13.47 h for T. castaneum, and 3.86 to 2.30 h for S. oryzae. It was found that S. oryzae adults were much more susceptible to the oil than T. castaneum. After 24 h of exposure, the LC₅₀ values (95% fiducial limit) for T. castaneum and S. oryzae were estimated to be 236.9 (186.27–292.81) and 3.34 (2.62–4.28) μl/l air, respectively. These results suggest that T. persicus essential oil merits further study as potential fumigant for the management of these stored-product insects.     

Aboveground biomass equations for individual trees of <Emphasis Type="Italic">Cryptomeria japonica</Emphasis>, <Emphasis Type="Italic">Chamaecyparis obtusa</Emphasis> and <Emphasis Type="Italic">Larix kaempferi</Emphasis> in Japan

Generic equations are proposed for stem, branch and foliage biomass of individual trees in even-aged pure stands of Cryptomeria japonica, Chamaecyparis obtusa and Larix kaempferi. Biomass data was collected from a total of 1,016 individual trees from 247 stands throughout Japan, and five regression models were assessed by root mean square error, mean bias, fit index (FI), and AIC. The results show that a power equation using diameter at breast height (dbh) and height is the most suitable for all species and components. This equation is more accurate than the familiar power equation that uses ‘dbh² height’, and it expresses the greater volume of branch and foliage mass of trees with a lower height/diameter ratio. A power equation using dbh is more reasonable for models with dbh as the only independent variable and more accurate than a power equation using ‘dbh² height’ for estimating branch and foliage mass. Estimating error for branch and foliage mass is larger than that for stem mass, but the entire aboveground biomass can be estimated with an error of less than 19%, except in the case of small trees with dbh less than 10 cm.     

A novel cold-inducible promoter,P<Emphasis Type="Italic">ThCAP</Emphasis> from <Emphasis Type="Italic">Tamarix hispida</Emphasis>, confers cold tolerance in transgenic <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>

Our previous studies have revealed that the ThCAP gene plays a vital role in transgenic Populus (P. davidiana × P. bolleana) in response to cold stress. However, the regulatory mechanism of ThCAP gene expression has been unclear. In this study, the 5′ flanking region of the ThCAP promoter (PThCAP) was cloned using a genome-walking method. By analyzing cis-acting regulatory elements of PThCAP, a DRE motif and MYC and MYB elements were found to be located in the promoter. To identify the regulatory elements that control the expression of the ThCAP gene promoter, a series of deletion derivatives of PThCAP, P1–P5, from the translation start code (?1538, ?1190, ?900, ?718 and ?375 bp), were fused to the GUS reporter gene, and then each deletion was stably introduced into Arabidopsis thaliana plants. Deletion analysis of the promoter suggested that only the P2 fragment had strong GUS expression in leaves and roots of A. thaliana exposed to low temperature stress. These results suggest that this 290-bp region (?1190 to ?900 bp), as an important part in PThCAP, was associated with cold tolerance of A. thaliana. Our results provide evidence for the regulatory mechanism of ThCAP gene involved in the response to cold stress, and that the gene is promising candidate gene for genetic improvement of crops.     

Structure and Function of <Emphasis Type="Italic">Populus deltoides</Emphasis> Agroforestry Systems in Eastern India: 1. <Emphasis Type="Italic">Dry matter dynamics</Emphasis>

Agroforestry systems based on poplar (Populus deltoides) are becoming popular in eastern and northern parts of India. Therefore studies on the structure and function of the systems are important. The investigations included allometric equations for above- and belowground tree components, crop and plantation floor biomass and litter fall estimation at Pusa, Bihar, India. Biomass, floor litter mass, litter fall and net primary productivity (NPP) of plantations increased with an increase in age of trees whereas, crop biomass for any specific crop interplanted with poplar decreased with the age of the plantation. The total plantation biomass increased from 12.08 to 90.59 Mg ha⁻¹ and NPP varied from 5.69 to 27.9 Mg ha⁻¹ year⁻¹. The biomass accumulation ratio ranged from 2.1 to 3.2. Total annual litter fall was in between 1.95 and 10.00 Mg ha⁻¹ year⁻¹, of which 92–94% was contributed by leaf litter. Compartmental models were developed for dry matter distribution in agroforestry systems involving young (3-year-old) and mature (9-year-old) poplar trees interplanted with various crops, the crops being grown in two rotations maize (Zea mays) – wheat (Triticum aestivum) – turmeric (Curcuma domestica) and pigeonpea (Cajanus cajan) – turmeric. This study substantiates the potential of Populus deltoides G₃ under agroforestry combinations.     

Potential contribution of <Emphasis Type="Italic">Quercus durata</Emphasis> and <Emphasis Type="Italic">Adenostoma fasciculatum</Emphasis> supplemented with <Emphasis Type="Italic">Medicago sativa</Emphasis> on intake and digestibility in sheep and goats

A digestion trial was conducted to estimate the potential contribution of the chaparral browse Quercus durata (leather oak) and Adenostoma fasciculatum (chamise) on intake and nutrient digestion in goats and sheep. Four wether Kiko goats (avg. wt. 22.9 kg) and four wether Targhee sheep (avg. wt. 39.6 kg) were housed in metabolism cages. Alfalfa pellets were used as a dietary supplement. Digestibility of the shrubs was measured by difference and metabolizable energy (ME) and ME intake (MEI) were calculated from digestibility and plants intake values. Oak and chamise had low crude protein (CP) content (6.5 and 4.3% DM) and high content of extractable condensed tannins (17.8 and 9.8% DM). In vitro organic matter digestibility (OMD) and ME for oak and chamise were 35.6 and 33.1% and 5.0 and 4.7 MJ/kg DM, respectively. In vivo OMD (calculated by difference with alfalfa) and MEI, for browse were greater (P < 0.01) for goats fed chamise compared to sheep. In oak, OMD was similar for both animal species but MEI was greater (P < 0.01) for goats. Greater (P < 0.05) DM and CP intakes (per kg BW^0.75) were observed for goats fed either oak or chamise compared with sheep. When an energy source was given as supplement, the potential contribution of chamise and oak to total ME requirements for maintenance was about 58.3 and 60.4% in goats and about 17.1 and 32.9% in sheep, respectively. In consequence, metabolizable energy supplied by these shrubs may not be adequate for maintenance when, during grazing, they are consumed in high proportions with other chaparral plants that may have lower nutritional value than the alfalfa supplemented in this study. Under an appropriate supplementation program, goats could be more effective than sheep to control fuel load in California chaparral, as they consume more chamise and leather oak and obtain more nutrients from them.     

<Emphasis Type="Italic">D</Emphasis><Superscript>2</Superscript> analysis for estimating genetic divergence in different clones of <Emphasis Type="Italic">Dalbergia sissoo</Emphasis>

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 D ² analysis revealed that D ² 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.     

Laboratory evaluation of the effectiveness of the entomopathogen; <Emphasis Type="Italic">Isaria farinosa,</Emphasis> on citrus mealybug, <Emphasis Type="Italic">Planococcus citri</Emphasis>

Citrus mealybug Planococcus citri (Risso) (Hemiptera: Pseudococcidae) is one of the main pests on citrus trees. Biological control of the pest is based on the release of hymenopterous parasitoids and coccinellid predators at present. The effectiveness of entomopathogen fungus Isaria farinosa (Holmsk.) Fries ([Sordariomycetes: Hypocreales] (Syn: Paecilomyces farinosus), as an alternative biological control agent on citrus mealybug, was investigated using four different inoculum densities and different relative humidities (RH). The entomopathogen caused 89.39% mortality in ovisacs, 84.07% mortality in second larval stage, 84.53% mortality in adult females, and 78.71% mortality in first larval stage at 95% RH and at 1 × 10⁸ conidia ml⁻¹ inoculum concentration. Percent mortalities were between 50 and 60 in ovisac, first and second larval stage at 95% RH and at 1 × 10⁷ conidia ml⁻¹. Percent mortality was decreased parallel to the decrease in humidity level and inoculum densities; however, the fungus caused significant infection in 70% and 80% RH. These results demonstrated that the effectiveness of the entomopathogen is promising for biocontrol of citrus mealybug.     

Effectiveness of Neem (<Emphasis Type="Italic">Azadirachta indica</Emphasis>) insecticides against Brassica pod midge (<Emphasis Type="Italic">Dasineura brassicae</Emphasis> Winn<Emphasis Type="Italic">.</Emphasis>)

The efficiency of the botanical insecticide (BI) NeemAzal T/S (containing 1% Azadirachtin A), on the basis of azadirachtin applied in a dose of 20 g a.i. ha⁻¹ against Brassica pod midge (Dasineura brassicae), has been monitored for 4 years. The biological efficiency of BI was compared with the efficiency of some synthetic insecticides. It was ascertained that BI was very efficient in decreasing the number of damaged oilseed rape pods (ranging from 56.5 to 85.9% compared to untreated plants) and its efficiency was comparable with synthetic insecticides based on Chloronicotinyl (Thiacloprid) and Neonicotinoid (Acetamiprid). BI’s efficiency was, in some years, even significantly higher compared to pyrethroid (λ-cyhalothrin). The high biological efficiency of azadirachtin that we ascertained was significantly reflected in increasing potential crop yields. The yield increase of azadirachtin ranged between 9.3 and 19.4% compared to the control sample. Azadirachtin showed the highest yield for the whole time of experimentation, and in some years the yield increase was significantly higher compared to some synthetic agents.     

Shoot multiplication of <Emphasis Type="Italic">Syringa reticulata</Emphasis> var. <Emphasis Type="Italic">mandshurica</Emphasis> from in vitro cultured seedlings

We developed a shoot multiplication protocol for Syringa reticulata Blume var. mandshurica Hara from in vitro cultured seedlings that derived from in vitro germinated seeds. The shoots could be induced on Murashige and Skoog (MS) medium with proper plant growth regulator combinations of 6-benzylaminopurine (BA) and indole-3-butyric acid (IBA). The better medium for shoot multiplication and growth was MS + 5 mg L^?1 BA + 0.5 mg L^?1 IBA + 20 g L^?1 sucrose + 7 g L^?1 agar, and the corresponding shoot induction rate was 75 %. The plantlets grew well after rooting on 1/2MS medium (macro-elements of MS medium are at half-strength) supplemented with 1 mg L^?1 IBA, and the survival percentage was >80 % at 16 weeks after transplanting.