Genetic diversity and relationship between cultivated clones of <Emphasis Type="Italic">Dalbergia sissoo</Emphasis> of wide geographical origin using RAPD markers

Random Amplified Polymorphic DNA (RAPD) polymor- phism was employed to assess the genetic diversity in the elite germ- plasm of Dalbergia sissoo. Sixty-seven clones that are under cultivation in northern India, originated from six different states of India and Nepal were analyzed with 30 RAPD primers that generated a total of 342 frag- ments out of which 290 were polymorphic. Total genetic diversity (Ht) varied between 0.01 and 0.37, with an average of 0.19. Shannon's In- formation index (I) varied between ...     

Growth and heritability estimates among clones of <Emphasis Type="Italic">Dalbergia sissoo</Emphasis> 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.     

Nursery practice on seed germination and seedling growth of <Emphasis Type="Italic">Dalbergia sissoo</Emphasis> 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·L⁻¹ respectively) 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.     

Identification of superior clones by RAPD technology in <Emphasis Type="Italic">Xanthoceras sorbifolia</Emphasis> Bge.

It is very important to select and identify superior Xanthoceras sorbifolia Bge.trees for purposes of breeding and exploration.For our study,we selected superior X.sorbifolia trees using RAPD (random amplification of polymorphic DNA) technology.The results show that certain specific amplification bands emerged with significantly different frequencies between superior and non-superior trees.The 1400-bp band of No.2121 primer emerged in the superior trees at a frequency 2.4 times that of the non-superior tree...     

<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.     

Effects of phosphorus and potassium addition on growth and nodula- tion 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.     

黑龙江省黑木耳9个栽培菌株遗传多样性的RAPD分析

本文对黑龙江省9个黑木耳 （Auricularia auricular）栽培菌株进行PCR-RAPD扩增和群体聚类分析, 在40个10bp随机引物中,筛选出13个引物.结果表明：黑木耳不同栽培菌株之间存在着丰富的多态性（96.05%）.用引物27和引物46的RAPD扩增指纹能有效分辨9个不同菌株的基因型,这一结果有助于在早期对黑木耳栽培菌株进行快速鉴别,为规范鉴别食用菌菌种混乱的局面提供科学依据.     

Clonal propagation of <Emphasis Type="Italic">Dalbergia sissoo</Emphasis> Roxb. and associated metabolic changes during adventitious root primordium development

A study was conducted on the twelve clones of shisham (Dalbergia sissoo Roxb.). These clones were obtained from India and Nepal. Single-node leafy cuttings were prepared from the vegetative multiplication garden to examine the clonal variation, effect of IBA treatment on rooting response and associated metabolic changes during adventitious root formation. A remarkable and significant variation was observed due to treatment of 2,000 ppm IBA in the rooting parameters. Clonal variations were also significant for root and shoot growth while length of root was insignificant. Among the twelve clones studied; C3 (Tulsipur, Gonda, Uttar Pradesh, India) and C4 (Laxmipur, Gonda, Uttar Pradesh, India) clone cuttings have given the highest rooting response. Interaction (clone × IBA) was significant only for production of number of roots per cutting. Periodic sampling for clone C3 was performed at 0, 7, 14, 21, 28 and 35 days to examine the contents of total soluble sugars, starch, protein and peroxidase (PER) activity in the rooting zone of cuttings (∼0.5 cm) during adventitious root primordium development. A significant increase in all the metabolic activities was noted due to IBA. Total soluble sugars and starch contents of cuttings decreased with the passage of time. Protein content and PER-activity started to increase in the early stage and reached the highest level on day 21, followed by a decline at the 35th day of sampling. These trends were common for both IBA treated and untreated cuttings. Protein content and PER-activity remained higher in the rooting zone of IBA treated cuttings. Overall these findings suggested that exogenous application of IBA may have activated carbohydrate metabolism for release of energy, while protein and PER-activity were necessary for cell division and differentiation during adventitious root primordium initiation and development in the rooting zone of cuttings.     

Growth,chlorophyll fluorescence and biochemical markers in clonal ramets of shisham (<Emphasis Type="Italic">Dalbergia sissoo</Emphasis> Roxb.) at nursery stage

About 19 clones of shisham (Dalbergia sissoo Roxb.) were raised by using healthy coppice shoot cuttings. After 2 year of establishment and growth, they were examined to identify suitable clone of shisham at nursery stage. Clones were studied for growth (height and basal diameter), chlorophyll fluorescence (Fv/Fm) and biochemical contents (chlorophyll a, b, total chlorophyll, free amino acids, total soluble proteins, total soluble sugars, starch and reducing sugar) in the leaves. Clone C2 and C3 (Gonda, Uttar Pradesh, India) had the highest growth and the chlorophyll fluorescence rate among examined clones. Between these two clones, C2 gave the highest response on height, basal diameter and Fv/Fm. Except free amino acids, all other biochemical contents were greater in the clones belonging to Gonda, Uttar Pradesh, India. Growth parameters, i.e. height and basal diameter increment at nursery stage had a positive correlation with all the studied parameters, except free amino acid and total soluble protein. These findings have demonstrated that there are remarkable clonal variations in growth, physiological and biochemical responses at nursery stage. Thus, physiological and biochemical markers can enhance selection efficiency, in addition to growth characteristics such as height and basal diameter as these have practical value in the tree improvement. The study suggests that clones of Gonda, Uttar Pradesh, India had the best performance and could be further tested in plantation programs.     

Comparison and early selection of new clones in <Emphasis Type="Italic">Populus tomentosa</Emphasis>

In our study, two experimental plantations, respectively, with 24 and 32 new clones of P tomentosa, were established in Weixian County, Hebei Province and Wuzhi County, Henan Province using a completely randomized block design. A comparative study was conducted on the continuous 5-year-old height and diameter at breast height （DBH） of new clones in the two plantations. As well, based on genetic correlation over the years of testing of these clones, a preliminary study of early selection was carried out. Results indicate that the growth traits of the new clones in Weixian were better than those in Wuzhi. The traits show weak correlation between the two plantations. In some stands, the height, DBH and seedling volume of 5-year-old clones presented statistically significant differences among clones. In both plantations, the new clones showed over 0.6 repeatability of height, DBH and volume, as well as larger coefficients of variation （CV）. The fact that these clones achieved the largest repeatability and CV in the second year suggests that these traits are highly controlled by heredity. Thus, based on the growth traits of the second year, the new clones B305, B307, B303, H75, BT18, BT17 and 21J-1 were considered suitable in Weixian. In Wuzhi, the new clones had variable repeatability and CVs in various years and their correlation of growth traits among different years was not high. We conclude that early selection of new clones was not feasible in Wuzhi.     

Small-scale genetic diversity in oneseed hawthorn (<Emphasis Type="Italic">Crataegus monogyna</Emphasis> Jacq.)

We have examined the pattern of random amplified polymorphic DNA (RAPD) variation among six populations of Crataegus monogyna from northern Italy, extending over an area of about 20,000 km². The aim was to evaluate local differentiation in relation to geographical distance. Of the 73 loci analysed, 65 were polymorphic, and there were no bands at complete fixation at the population level. The mean genetic diversity was 0.291, with the values for individual populations ranging between 0.252 and 0.333. According to the analysis of molecular variance most of the genetic variation was found within populations (about 80%), with a significant proportion being attributable to genetic differences between populations. No evidence for isolation by distance was found in the set of populations sampled. The efficacy of RAPD markers in analysing genetic variation, and the contribution of the results in terms of the preservation of biodiversity and defining the most appropriate strategies for collecting forest reproductive material, are discussed.     

Assessment of morphological and genetic diversity in <Emphasis Type="Italic">Gmelina arborea</Emphasis> Roxb.

Gmelina arborea is an important timber-yielding tree that grows naturally in the tropical and subtropical regions of Southeast Asia and has also been introduced as a plantation species outside these regions. Genetic diversity in this tree species was observed in stone/seed-related traits and in vitro responses of cultured nodal segments from plants of eight different populations representing natural forests, fragmented forests and plantations. Variance analysis showed significant differences between populations for these traits. However, it was not possible to separate the different populations using these traits by multivariate analysis, even after environmental variation was reduced over six subcultures. Genetic diversity was therefore analysed using molecular markers. Inter-simple sequence repeat (ISSR) primers yielded 95% polymorphic loci among the eight populations and UPGMA analysis enabled separation of these populations on the basis of their genetic distances. Diversity was also analyzed using population genetics parameters like Nei’s genetic diversity and gene differentiation. Nei’s genetic diversity was 0.29 between populations and 0.11 within populations. AMOVA analysis indicated 41 and 59% within- and between-population genetic diversity, respectively. Mantel test suggested that genetic differentiation between six Indian populations was positively correlated to geographic distance (r = 0.626, P = 0.029). Assessment of the genetic variation in G. arborea populations is an important step in selection of conservation strategies for this species since diversity forms the basis for species adaptation.     

Feeding performance of <Emphasis Type="Italic">Clostera fulgurita</Emphasis> on three clones of <Emphasis Type="Italic">Populus deltoides</Emphasis>

Poplar leaf defoliator, Clostera fulgurita (Walker) larvae were reared on three Populus deltoides clones (PL1, PL5 and PL7) in the laboratory. The nutritional indices were computed for working out the relationship between food consumption and growth rate of 3rd, 4th and 5th instar larvae on three clones. The result showed that the consumption index (CI), approximate digestibility (AD), growth rate (GR), relative growth rate (RGR) and efficiency of conversion of ingested food (ECI) decreased with the increase in the age of the larvae. Efficiency of conversion of digested food (ECD) increased with increase in age of the larvae. GR and RGR varied significantly, indicating that larval development was enhanced on PL1 as compared to PL5 & PL7. The values of AD, ECI and ECD were not affected by the different clones. Feeding and growth indices could be useful to define a defoliation prediction model.     

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.     

Population genetic structure and diversity of high value vulnerable medicinal plant <Emphasis Type="Italic">Acorus calamus</Emphasis> in India using RAPD and chloroplast microsatellite markers

Acorus calamus is a highly valued medicinal plant with global distribution used in several drugs of health care systems. We evaluated the genetic diversity and population structure of 50 populations of A. calamus from different geographical regions in India through RAPD and chloroplast microsatellite markers. From the total screened 82 RAPD primers and 18 cpSSR primers, 10 RAPD and nine cpSSRs were found polymorphic. The selected 10 RAPD primers produced a total of 96 reproducible bands, out of which 65 wer...     

冷湿层积法对印度冷杉(Abies pindrow)和长叶云杉(Picea smithiana）种子萌发的影响

研究赤霉素(GA3),冷湿和温度 对五个种源的印度冷杉(Abies pindrow)和长叶云杉(Picea smithiana)种子萌发的影响.种子被浸泡在GA3 (10 mg(L-1)中24小时,然后在3(5(C温度的条件下冷藏15天.设计4个温度(10(C, 15(C, 20(C 和 25(C)条件来促进种子的萌发.结果表明,浸泡和冷湿处理明显增加种子的萌发率.在10 (C时种子的萌发率最高.总体结果表明,种子被浸泡在GA3 (10 mg(L-1)中24小时,冷湿藏15天,可以有效地促进印度冷杉和长叶云杉的种子萌发.统计数据表明,浸泡处理、温度和种源以及与温度的相互作用都对种子的萌发有明显的影响.     

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.     

Realized genetic gain observed in second-generation seedling seed orchards of <Emphasis Type="Italic">Acacia mangium</Emphasis> in South Kalimantan,Indonesia

Realized genetic gains brought by the first-generation tree improvement of Acacia mangium were evaluated by analyzing the 1-year growth performance of growth (height, dbh) and form (stem straightness, multi-stem) traits using data collected from three second-generation orchards: groups B, C, and D. Each orchard was composed of improved families derived from open-pollinated seed of plus trees selected from the first-generation seed orchards and newly added unimproved families collected from the same geographic region. The number of improved families and that of the unimproved in each orchard were 31 and 23 for group B, 28 and 39 for group C, and 24 and 30 for group D. Gains were calculated as percentage increase of improved families over unimproved ones. Improved families performed better than unimproved ones in 11 out of 12 comparison cases: four traits each in the three orchards. Averages of the realized genetic gain across the three seedling seed orchards were 3.1%, 5.2%, 4.3%, and 0.5% for height, dbh, stem straightness, and multi-stem, respectively. Except for multi-stem, the performances of improved families were significantly better than those of unimproved families in all the three orchards for dbh and in two out of three orchards for height and stem straightness.     

Nutrient dynamics associated with leaf litter decomposition of three agroforestry tree species (<Emphasis Type="Italic">Azadirachta indica,Dalbergia sissoo</Emphasis>, and <Emphasis Type="Italic">Melia azedarach</Emphasis>) of Bangladesh

Azadirachta indica A. Juss, Dalbergia sissoo Roxb., and Melia azedarach L. are little studied species in nutrient return capabili- ties from leaf litter decomposition to maintenance of the soil fertility despite their importance in agroforestry practices of Bangladesh. A leaf litter decomposition experiment was conducted using a litterbag tech- nique to assess the nutrient return efficiency of these species. The de- composition rate of leaf litter was highest for M. azedarach and lowest for D. sissoo. Rainfall and temperature of study sites showed a signifi- cant (p<0.05) positive relationship with the rate of leaf litter decomposi- tion. The highest decay constant was observed for M. azedarach (6.67). Nitrogen and Phosphorus concentration in leaf litter showed a decreased trend sharply at the end of the first month, whereas rapid decrease of Potassium concentration was reported within 10 days. Conversely, higher concentration of nutrient was observed at the later stages of decomposi- tion. All three species showed a similar pattern of nutrient release (K > N > P) during the decomposition process of leaf litter. Among the studied species, D. sissoo was best in terms of N and P return and A. indica was best in terms of K return.     

Molecular identification of sex in <Emphasis Type="Italic">Hippophae rhamnoides</Emphasis> L. using isozyme and RAPD markers

In many dioecious plants, gender affects economic value, breeding schemes and opportunities for commercial harvests. Hippophae rhamnoides L. is a dioecious plant species in which female genotypes are commercially preferred over male genotypes. Its berries have rich medicinal, nutritional and pharmaceutical properties because of their large amounts of vitamins, essential oils, proteins, fatty acids, free amino acids and flavanoids. Primary limitation for breeding H. rhamnoides L. is its dioecious nature, since gender cannot be identified by traditional methods. Therefore, some reliable and quick methods need to be developed. This communication deals with the development of isozyme and RAPD markers for early sex identification in this dioecious tree. The isozyme analysis was conducted with four enzyme systems, viz. peroxidase, esterase, malate dehydrogenase and catalase. The peroxidase enzyme system produced a female specific sex marker, which successfully differentiated between the staminate and pistillate genotypes of H. rhamnoides L. Thirty five random decamer primers were used in our study and one male sex linked marker was identified. OPD-20 (5′-ACTTCGCCAC-3′) displayed a band at 911 bp that expressed polymorphism between male and female genotypes. The staminate and pistillate genotypes could be distinguished using RAPD marker OPD-20911. These results revealed the immense potential of peroxidase isozyme patterns and RAPD as genetic markers for sex identification in H. rhamnoides L.