Factors influencing in vivo and in vitro micrografting of sandalwood (Santalum album L.): an endangered tree species

In vivo micrografting of Santalum album was achieved (50%) by grafting 4- to 5-cm-long scions, collected from a candidate plus tree (CPT) of 50–60 years of age, onto 90-day-old nursery-grown rootstock. Scion size, rootstock age, and scion collection season were found to influence graft success. Grafted plants were incubated under greenhouse conditions for 6–8 weeks during the graft union process. In vitro micrografts were achieved by placing 1- to 2-cm-long scions derived from nodal shoot segments (collected from CPT) onto the hypocotyl of 45-day-old in vitro rootstocks. Use of in vitro grown shoots as a source of scion gave better graft success (60%) than scions collected directly from field-grown trees. In vitro grafting was also influenced by scion size and rootstock age. Under favorable conditions, scions and hypocotyls unite to form complete plants that produced two to four leaves after 6–8 weeks. This is the first report on in vivo and in vitro micrografting of S. album having potential for production of disease-free clonal plants for conservation and improvement targets.     

Effects of different culture media on the growth of Indian sandalwood （Santalum album L.） seedlings in Zhanjiang, Guangdong, southern China

We studied the effects of different culture media on the growth of India sandalwood(Santalum album L.) seedlings in Zhanjiang,Guangdong Province in southern China. Five different growth substrates,lateritic subsoil,burnt soil,agricultural soil,peaty soil and coconut dust,were used as the basic culture materials and seven different treatments of composition were used as potting media. Kuhnia rosmarinifolia Vent. was used as a primary host plant for all treatments. Statistically significant differences were f...     

Effects of different culture media on the growth of Indian sandalwood (Santalum album L.) seedlings in Zhanjiang, Guangdong, southern China

We studied the effects of different culture media on the growth of India sandalwood (Santalum album L.) seedlings in Zhanjiang, Guangdong Province in southern China. Five different growth substrates, lateritic subsoil, burnt soil, agricultural soil, peaty soil and coconut dust, were used as the basic culture materials and seven different treatments of composition were used as potting media. Kuhnia rosmarinifolia Vent. was used as a primary host plant for all treatments. Statistically significant differences were found between treatments in respects of survival rate (p < 0.001), height (p < 0.001), ground diameter (p < 0.001) and biomass (p = 0.002), as well as for quality index (p = 0.001) of S. album seedlings after 6-month growth in containers with different culture media. Among all treatments, the treatment combining burnt soil, peat and coconut dust in a weight ratio of 1:1:1 plus 2% calcium super-phosphate as basal manure achieved the best performance for most of the seedling growth parameters, including survival rate (98%), height (35.81 cm), ground diameter (0.56 cm), biomass (4.46 g) and quality index (0.65), followed by the treatment using only burnt soil plus 2% calcium super-phosphate as the culture medium (survival rate 86%, height 29.23 cm, ground diameter 0.48 cm, biomass 3.36 g and quality index 0.52), while the treatment using only lateritic subsoil plus the basal manure as the medium obtained the poorest results in survival rate (38%), height (12.04 cm), ground diameter (0.19 cm), biomass (0.26 g) and quality index (0.043). Increasing the proportion of lateritic subsoil in the medium when mixed with peat and coconut dust did not show statistically significant differences in survival, height, ground diameter, biomass nor in quality index. In consideration of cost, using burnt soil plus 2% calcium super-phosphate as basal manure may be the optimum culture medium for large-scale production of Indian sandalwood seedlings in Guangdong, southern China.     

Micropropagation of Vitex negundo L. and Random Amplified Polymorphic DNA Analysis of Micropropagated Plants

A micropropagation protocol was established for a medicinal plant Vitex negundo. Genetic stability of micropropagated plants was investigated. Multiple shoots were induced from nodal explants cultured on Murashige and Skoog (MS) medium with 0.53 μM naphthalene acetic acid (NAA) and 11.0 μM benzyl aminopurine (BAP) along with additives (ascorbic acid, 283.9 μM; citric acid, 130.1 μM; and arginine, 143.6 μM). Shoots were further multiplied by repeated transfer of the mother explant. The shoots were further multiplied on MS medium + 0.57 μM indole-3-acetic acid (IAA) and 6.6 μM BAP. The micropropagated shoots were pulse treated with 122.5 μM indole-3-butyric acid (IBA), in liquid MS medium and then transferred to autoclaved soilrite. These rooted ex vitro. Shoots were also rooted in vitro on a half-strength MS medium + 2.45 μM IBA. The survival rate of in vitro rooted plantlets was poor during hardening compared to ex vitro rooted plantlets. About 95% of the ex vitro rooted, hardened plantlets survived in the field. Genetic stability of micropropagated plants was tested by using 25 random amplified polymorphic DNA primers. The cloned plants exhibited no variation in banding pattern in comparison with the mother plant.     

巨桉组织培养及工厂化育苗技术的研究

巨桉是造纸和人造板的良好材料,其组织培养微繁殖体系已经研究成功。取优树主干基部萌芽或扦插苗1~1.5 cm长的茎段做为外植体,外植体和增殖培养在附加有BAP和NAA的MW培养基上进行。培养条件为26±2℃,日光灯光照10 h.d-1。增殖培养采用BAP 0.1~0.5 mg.L-1和NAA 0.01~0.1 mg.L-1。离体的茎芽适合在B2附加0.5 mg.L-1IBA生根。小苗移栽红心土上,在简易温室成活率高达93%。     

Desmanthus virgatus (L.) Willd. An Efficient Intermediate Host for the Parasitic Species Santalum album L. in Timor,Indonesia

In cultivation, sandalwoods suffer high seedling losses both in the nursery and when transplanted to the field. It was hypothesised that early haustorial connection to an efficient pot host should enhance growth and survival in both nursery and field. Nursery performance of Santalum album was contrasted with a range of species as initial- or nursery-stage hosts for plantation establishment in Timor. With pot-hosts of Desmanthus virgatus, Crotalaria juncea L. or Alternanthera cv., sandalwood grows rapidly to a mean height > 25 cm at < 26 weeks from sowing. These species are superior pot-hosts to the traditional Capsicum frutescens L. Subsequent field performance over 4 years is best with those sandalwoods initially hosted to D. virgatus. This species has the capacity to function as both a pot-host and an intermediate field-host for sandalwood. We conclude that for Timor conditions, sandalwood should be preferentially grown with D. virgatus as the first-stage, or in-pot, host plant species.     

In vitro clonal propagation of mature Tectona grandis through axillary bud proliferation

An in vitro clonal propagation procedure for mature Tectona grandis (teak) trees is described. Multiple shoots were induced from nodal segments through axillary bud proliferation. A shoot multiplication rate of 6.33 was achieved on Murashige and Skoog's (MS) medium supplemented with 10 μM 6-benzyladenine BA and 1 μM 1-naphthalene acetic acid (NAA) during every subculture cycle of 4 weeks. In vitro raised shoots could be successfully rooted (66.66%) on liquid MS medium supplemented with 15 μM NAA, with 1.60 roots per shoot, every 6 weeks of culture. In vitro hardening was carried out in sand soaked with half-strength MS medium (organic free). The plantlets were acclimatized first in a mist chamber and then in polybags in a mixture of soil, sand, and farmyard manure (1 : 1 : 1 v/v) in a shade house.     

Preservation of Juniperus thurifera L.:a rare endangered species in Algeria through in vitro regeneration

Juniperus thurifera L.is an endemic Cupres saceae from the Aure`s Mountains of north eastern Algeria and endangered,in part,due to the scarcity of viable seeds It is threatened by other abiotic factors and the lack of an effective management strategy will increase its risk o extinction.The dearth of information on its in vitro regeneration impedes its application in forest managemen programs.We therefore developed a micropropagation protocol using microcuttings with auxiliary buds.Cuttings were grown on different combinations of media supplemented with plant growth regulators at different concentrations.The highest number of shoots and branches regenerated from original shoots was obtained on Woody Plant Medium(WPM)supplemented with 6-benzylaminopurine(BAP)(0.5 mg L^-1)and 2,4-dichlorophe noxyacetic acid(2,4-D)(0.25 mg L^-1).The best elongation of shoots was achieved with WPM supplemented with0.5 mg L^-1of BAP and 0.25 or 1 mg L^-1 of 2,4-D.On the second subculture,shoots had a higher number of branches than those of the first.The highest rooting rate,38.8%,was obtained with shoots cultured in 1/2 Murashige and Skoog(MS)medium supplemented with 5.0 mg L^-1each of indol-3-butyric(IBA)and naphthalene acetic acid(NAA).Similarly,the highest root numbers and lengths were produced on 1/2 MS medium supplemented with IBA and NAA(5.0 mg L^-1each).During transfer to acclimatization,rates of plant losses of 50% occurred.The second part of the experiment showed that the best shoot callusing was on WPM supplemented with BAP and 2,4-D,with either the combination 0.5+0.25 or 0.25+0.25 mg L^-1.The results of this research provide a starting point for further studies on in vitro regeneration of J.thurifera for the sustainable management of its unique ecosystem in the Mediterranean basin.     

无花果品种“中国紫果”离体培养与保存研究

本研究对无花果品种“中国紫果”的离体培养再生体系进行了探讨。结果表明,茎尖增殖的培养基为MS＋6-BA0.5mg／L＋NAA0.05mg／L,生根培养基为1／2MS＋IBA 0.2mg／L。将生根的无菌苗进行驯化后移栽到营养钵中,成活率可达100％。试验还表明GA3对“中国紫果”组培苗茎的伸长生长有显著效应。另外,试验探讨了“中国紫果”离体种质保存的限制生长保存方法,结果发现在1／2MS培养基（1．5％蔗糖）中,新梢基本无生长,7个月后转入含0．2mg／LGA3的增殖培养基中能迅速恢复生长。可见,用此方法可达到较好的短期保存效果。     

Micropropagation of Leptadenia pyrotechnica (Forsk.) Decne: A Multipurpose Plant of an Arid Environment

Leptadenia pyrotechnica (Forsk.) Decne belongs to the family Asclepiadaceae. It is commonly known as Kheemp in India. L. pyrotechnica is an important component of an arid ecosystem and source of fiber, forage, and medicines. We report here in vitro propagation protocol of L. pyrotechnica. Cotyledonary nodes of in vitro raised seedlings were used as an explant for multiplication of shoots. Shoots were multiplied on Modified Murashige and Skoog's (MMS) medium containing 1.33 μM BAP and additives (50 mg L^?1 ascorbic acid and 25 mg L^?1 each of citric acid, arginine, and adenine). Cultures were maintained at 30 ± 2°C temperature, 50–60 μmol m^?2 s^?1 SFP, 12 hr day^?1 photoperiod, and 60% relative humidity (RH). In vitro multiplied shoots were rooted in vitro on half-strength MS medium containing 2.46 μM IBA and 100 mg L^?1 activated charcoal. Shoots were rooted ex vitro using 2460 μM IBA pretreatment for 15 min. Plantlets were hardened in a greenhouse, and survived on a mixture of sand, garden soil, and organic manure in 3:1:1 ratio in polybags in natural habitats.     

光叶楮组织培养快速繁殖技术的研究

对光叶楮组织培养中的外植体灭菌、苗玻璃化预防、增殖培养、根原基诱导及室外移栽进行的试验结果表明:以优良母株的具腋芽茎段为外植体,经过0.1%的HgC l2灭菌后,外植体成活率为53.6%;培养基中添加4.0%的蔗糖及增加培养容器的透气性能能较好地预防玻璃苗的发生;在MS培养基中附加BA,NAA等植物生长调节剂可有效促进试管苗的增殖和生长,其中以1.5 mg/L BA 0.1 5 mg/L NAA的组合最为适宜;适宜的根原基诱导培养基为1/2MS 0.5 mg/L IBA 0.3 mg/L NAA 100 mg/L LH 2%蔗糖;具根原基试管苗移栽于混合基质(蛭石∶珍珠岩∶泥炭=6∶3∶1,体积比)中,温度控制在20~30℃,相对湿度为85%~90%,保湿15~20 d,其间适当遮荫,30 d后试管苗成活率达90%。     

Competition for light, water and nitrogen in an association of hazel (Corylus avellana L.) and cocksfoot (Dactylis glomerata L.)

Competition for light, water and nitrogen between hazel trees and cocksfoot grown under the trees was studied in two hazel plantations managed in different ways. The first plantation with a light transmission of at least 70% at grass level was compared with two control hazel and cocksfoot monocrops. The soil was a calcareous heavy clay, 1.4 m deep. Water stress during summer was severe in the intercropped stand as well as the grass- only plots, while it was much less in the sole stand of trees (without grass). Hazel tree water potential in the intercropped stand improved in the fourth year as the trees extended their roots similarly to the grass-free ones, despite their much lower canopy growth rate. The fourth year, local N fertilisation began to be effective as evidenced by the N content per leaf area unit and growth of the intercropped trees: grass root competition was high and root barriers were needed to exclude it. The second plantation was managed on a deep soil with a water table at approximately 2.5 m depth. The trees did not suffer from the presence of grass, and allowed a 15 to 75% light transmission rate. A control monocropped grass plot was established for comparison. On the basis of the multiple limitation hypothesis, a grass dry matter production model was built which accounts for radiation transmitted to the understorey, water supply, temperature and air moisture deficit in interaction with the radiation transmission rate (r2 = 0.716; 590 observations). The limits of such models are discussed with regard to insufficient knowledge on root dynamics. This revised version was published online in June 2006 with corrections to the Cover Date.     

Impact of artificial pruning on growth and secondary shoot development of wild cherry (Prunus avium L.)

Producing high value veneer wood requires that the tree bole be branch-free. This can be accomplished by natural or artificial pruning. Since wild cherry does not self prune well, pruning artificially is the only practical option. The study analysed the effect of conventional whorl-wise pruning and selective pruning, on height growth, diameter growth and secondary shoot development of wild cherry. Four pruning treatments were applied on cherry trees in summer 2007, one group of cherries was left unpruned to serve as a control: treatment C1 (upper 5 whorls left), C2 (upper 3 whorls left), S1 (removal of branches larger than 3 cm or with an angle to the stem < 40°), S2 (removal of branches larger than 2 cm or with an angle to the stem < 40°), N (unpruned). Data showed that height growth was not affected by pruning. In contrast, diameter growth at breast height of the C2 pruned cherry was reduced by approximately 5% (SE = 2.7%) in the year of pruning (trees were pruned in July). This pruning treatment produced significant (p = 0.028) nine percent less diameter growth than the control in the second year following pruning. The diameter increment of the C1 pruned trees with five whorls left after pruning and the selective pruned cherries were only about 4% (SE = 4.0%) smaller than the control after two years. This loss was statistically not significant. Analyses showed that on selective pruned trees the survival rate of secondary shoots was significantly reduced compared to those on whorl-wise pruned trees. Significant differences in the size of the secondary shoots were only found between the C1 and S1 (p < 0.05) pruned trees. We did not find differences in the total number of secondary shoots per tree among pruning treatments. Solely from a tree growth perspective, the moderate whorl-wise pruning treatment C1 and the selective prunings were equally effective in minimizing the reduction of diameter growth and are recommended in practice. However it was found that the survival of secondary shoots was reduced on selective pruned trees although the amount of pruning work needed in selective pruning was slightly greater than conventional moderate pruning.     

Early growth and seasonal chemical composition of three indigenous multipurpose tree species (MPTS) in Abeokuta,Nigeria

A study was carried out to evaluate the growth parameters and nutritive qualities of Moringa oleifera, Millettia griffoniana and Pterocarpus santalinoides. The main objective of the study was to assess the potential of the MPTS in supplementing the feed of ruminant animals during the dry season when grasses are scarce and their quality generally fall short of animal requirements. Leaf samples were randomly collected from the trees for estimation of the proximate composition, neutral detergent fibre, acid detergent fibre and gas production using the in vitro technique. Samples were collected three times to represent seasonal variations as follows: November: Early Dry; February: Mid Dry and April: Late Dry seasons. Gas production was recorded at 3, 6, 9, 12, 15, 18, 21, 24, 48, 72 and 96 h of incubation. Mortality rate was less than 5%. M. oleifera recorded the best overall growth respectively followed by P. santalinoides and M. griffoniana. P. santalinoides and M. oleifera retained more leaves in the dry season than M. griffoniana. The dry matter (DM) and ether extract (EE) contents of the MPTS were significantly (P < 0.05) affected by seasons and plant species while there were significant (P < 0.05) species effects on the CP and ADF values. The gas production values and the fermentation parameters indicated the presence of potentially degradable nutrients in the MPTS, which underscores the importance of the tree species as sources of nutrient for ruminant animals during the dry season.     

The process of decline of an endangered tree species,Pinus armandii Franch. var.amamiana (Koidz.) Hatusima, on the southern slope of Mt. Hasa-dake in Yaku-shima Island

The process of decline of an endangered tree species,Pinus armandii var.amamiana, was monitored on the southern slope of Mt. Hasa-dake in Yaku-shima Island from 1994 to 1998. There are 163 trees ofP. armandii var.amamiana over 1.3 m in height. They are distributed on steep ridges and rocks with a thin soil layer mostly consisting of friable granite. During the monitoring period, 21 trees ofP. armandii var.amamiana died and the mortality rate was 12.9%. Dead trees were categorized into three types: standing, uprooted and landslide. The uprooted- and landslide-typed dead trees were found only after a severe typhoon struck Yaku-shima Island. This suggests that the combination of fragile site conditions and severe typhoons play an important role in the process of decline ofP. armandii var.amamiana. The standing-typed dead trees were presumed to have been killed by pine wilt disease, accounting for 71.4% of the dead trees. However,Bursaphelenchus xylophilus, the cause of pine wilt disease, was not detected from any of the wood chips or branch samples from the standing-typed dead trees ofP. armandii var.amamiana. This might indicate that some factor(s) other than pine wilt disease could be responsible for the standing-typed death ofP. armandii var.amamiana in natural habitats. A part of this paper was presented at the 109th Meeting of the Japanese Forestry Society.