In Vitro Propagation and Artificial Seed Production of Fritillaria cirrhosa D. Don, an Endangered Medicinal Plant

Fritillaria cirrhosa D.Don(Liliaceae)is an endangered perennial bulbous plant and its dry bulb is a valuable med-icinal material with antitussive and expectorant effects.Nevertheless,lack of resources and expensive prices make it difficult to meet clinical needs.This study presents a regeneration system aimed at overcoming the challenge of inadequate supply in F.cirrhosa,focusing on:(1)callus induction,(2)bulblets and adventitious bud induction,and(3)artificial seed production.Callus development was achieved in 84.93%on Murashige and Skoog(MS)medium fortified with 1.0 mg·L^(-1) picloram.The optimal medium for callus differentiation into regenerated bulb-lets was MS medium supplemented with 1.0 mg·L^(-1)6-benzyladenine(6-BA)and 0.2 mg·L^(-1)α-naphthaleneacetic acid(NAA).Subsequently,bulblets and adventitious buds were induced from regenerated bulblet sections cul-tured on MS medium fortified with 0.3 mg·L^(-1)6-BA+1.0 mg·L^(-1)2,4-dichlorophenoxyacetic acid(2,4-D),2.0 mg·L^(-1)6-BA+0.5 mg·L^(-1),and the induction rates were 88.17%and 84.24%,respectively.The regenerated bulblets were transplanted into a substrate of humus soil,river sand,and pearlite(1:1:1)after low-temperature treatment.The germination rate was 42.80%after culture for 30 days.Regenerated bulblets were used for encap-sulations in liquid MS medium containing 3%sucrose(w/v)+0.5 mg·L^(-1) NAA+2.0 mg·L^(-1)6-BA+3%sodium alginate(w/v)with a 10 min exposure to 2%CaCl_(2).Under non-aseptic conditions,the germination rate reached 81.67%,while the rooting rate was 20.56%after 45 days.The capsule added 1.0 g·L^(-1) carbendazim and 1.0 g·L^(-1) activated carbon was the best component of artificial seeds.This study successfully established an efficient regen-eration system for the rapid propagation of F.cirrhosa,involving in vitro bulblet regeneration and artificial seed production.This method introduces a novel approach for efficient breeding and germplasm preservation,making it suitable for large-scale industrial resource production.

Effects of Different Seasons on Cutting Propagation of Five Citrus Rootstocks

[Objectives]To explore the cutting propagation mode of citrus rootstock,improve the survival rooting rate of citrus rootstock,and provide theoretical guidance and technical reference for the rooting research of difficult-to-root plants such as citrus.[Methods]Five citrus rootstocks Citrus tangerina Tanaka‘Hongju’,Citrus haniana Hort‘Suanju’,Citrus limonia Osbeck‘Hongningmeng’,Citrus sinensis×Poncirus trifoliata‘Zhicheng’and Poncirus trifoliate(L)Raf.‘Zhike’were used as experimental materials to select the suitable cutting substrate for citrus rootstocks by measuring the physical properties of the substrate.Cutting was carried out in spring,summer,autumn and winter respectively.After cutting,the morphological changes of cuttings were observed regularly,and the callus rate,germination rate and rooting rate of cuttings were recorded.[Results]The best substrate for citrus rootstock cutting was peat soil,vermiculite and fine river sand(2:1:1).The callus of citrus rootstock in different cutting seasons began to appear in 10-22 d,and the callus rate reached 55%-100%.In terms of budding,the budding time was the earliest in summer and autumn,and slightly later in spring and winter;the germination rate of C.limonia Osbeck‘Hongningmeng’was the highest,and the germination rate of C.sinensis×P.trifoliata‘Zhicheng’and P.trifoliate(L)Raf.‘Zhike’was lower;in terms of rooting,C.limonia Osbeck‘Hongningmeng’had the earliest rooting time and the highest rooting rate and could reach 100%in all seasons;the rooting rate of C.tangerina Tanaka‘Hongju’was 50%-80%;the rooting rate of C.haniana Hort‘Suanju’was 60%-80%;C.sinensis×P.trifoliata‘Zhicheng’and P.trifoliate(L)Raf.‘Zhike’showed the earliest rooting time and the highest rooting rate in summer,and the latest rooting time and the lowest rooting rate in winter,which were only 14.5%.Therefore,different citrus rootstock varieties should choose the appropriate cutting time according to their own characteristics.[Conclusions]The results of this study can provide a scientific basis for a large number of cutting propagation of different citrus rootstocks,and have practical guiding significance for large-scale planting.

Aseptic Seedling and Rapid Propagation of Ludisia discolor

[Objective] This study was conducted to establish a system for aseptic seeding and rapid propagation of a wild Chinese herbal medicine Ludisia discolor. [Method] The seeds of L. discolor were germinated in four different culture media MS, 1/2MS, Hyponex No.1 （3.0 g/L）, and 1/2 Hyponex No.1 （1.5 g/L）, and the ger- mination rate was calculated 80 d later to select the optimal medium for seed ger- mination of L. discolor. Then, by culturing the protocorms and buds of L. discolor in the media supplemented with different hormone combinations （6-BA and NAA） or organic substrates （potato juice, banana juice and coconut milk）, the optimal medi- um compositions for multiplication and .differentiation, seedling hardening and rooting of L. discolor were respectively determined. [Result] After 80 d of dark culture, 86.67% of the seeds inoculated into the medium 1/2 Hyponex No.1 germinated and developed into protocorms, proving 1/2 Hyponex No.1 was most suitable for seed germination of L. discolor among the four basic media, while MS medium had the worst effects. In culture medium of 1/2 Hyponex No.1＋2.0 mg/L 6-BA＋100 ml/L co- conut milk supplemented with 0.4 or 0.6 mg/L NAA, a mixed body of protocorms and small buds were induced after being cultured for 45 d, and the multiplication rate was 6.11, which was higher than that in the medium supplied with 0.2 or 0.8 mg/L NAA. By adding 10% potato juice, 10% banana juice or 100 ml/L coconut milk into Hyponex No.1＋1.0 mg/L NAA, it was found that 10% banana juice had better effects in hardening and rooting of L. discolor planlets than 10% potato juice and 100 ml/L coconut milk. The small buds grew into plantlets about 80 d later in the medium with banana juice, the rooting rate was 93.33%, and the plantlet height was 7.19 cm. After hardening-culture for 10 d in a greenhouse, the L. discolor plantlets were transplanted into the mixed matrix of peat soil and lava （at a ratio of 3：1）, and 91.7% of them survived 60 d later. [Conclusion] A relatively perfect propa- gation system of L. discolor was established by using aseptic seeds as explants in this study, which can be used for the protection of L. discolor germplasm resources, seedling propagation and industrial development.

Single-seed sowing increased pod yield at a reduced seeding rate by improving root physiological state of Arachis hypogaea

Double-seed sowing(two seeds per hole)is the dominant pattern of peanut sowing in China,but within-hole plant competition usually limits their growth and yield formation.Besides,the traditional double-seed sowing method does not facilitate mechanization during sowing.The objective of this study was to determine if single-seed sowing at a proper seeding rate yielded better than traditional double-seed sowing pattern and the differences of physiological metabolism of roots.A field experiment was conducted in two consecutive years to compare pod yields of single-seed sowing at 180000(S180),225000(S225),and 270000 seeds ha^-1(S270)with that of double-seed sowing at 270000 seeds ha^-1(D270)using a completely randomized block design with four replications.And the root bleeding sap rate,nutrient content,and the main hormone contents in root bleeding sap were also comparatively investigated.Although the pod yields of single-seed sowing at the three densities were higher than that of traditional double-seed sowing(D270),S225 yielded better than the other two single-seed sowing treatments(S180 and S270).The increased pod yield in single-seed sowing at 225000 seeds ha^-1 was mainly due to the higher pod dry weight per plant and harvest index.The improved pod dry weight and shoot growth had closely relationship with the enhanced root physiological traits such as the increased root bleeding sap rate,content of free amino acids,soluble sugars,K^+,Mg^2+,Zn^2+,and Ca^2+of the individual plant root.The improved activity of root reductive,nitrate reductase(NR)and ATPase and higher zeatin and zeatin riboside(Z+ZR)content of root bleeding sap were alsocrucial to the pod and shoot growth of peanut.Single-seed sowing at a moderate seeding rate(S225)is a potential practice to increase pod yield and to save seed cost.

Lodging Resistance Related to Root Traits for Mechanized Wet-Seeding of Two Super Rice Cultivars

Mechanical hill wet-seeded rice machine is beneficial for establishing and growing uniform rows of seedlings.However,there is limited knowledge regarding the effects of the establishment of furrows on growth,lodging and yield,and their relationships with root traits.In this study,field experiments were conducted during 2012 and 2013 using two super rice varieties(hybrid rice Peizataifeng and inbred rice Yuxiangyouzhan)under three furrow establishment treatments(T1,both water and seed furrows were established by the machine;T2,only seed furrows were established by the machine;and T3,neither water nor seed furrows were established by the machine).Lodging index,lodging-related traits,grain yield,above-ground dry weight and root traits were measured.The results showed that the lodging index was significantly affected by the treatments with furrows(T1 and T2).The strongest lodging resistance was detected in the mechanical hill wet-seeded rice with furrow treatment(T1)in both 2012 and 2013.Lodging resistance was strongly related to the breaking resistance.No significant difference was found in grain yield or dry weight of the mechanical hill wet-seeded rice.Therefore,the mechanical hill wet-seeded rice with furrow treatment increased rice lodging resistance,which was related to root traits.

In vitro germination and propagation of a threatened medicinal orchid,Cymbidium aloifolium(L.) Sw.through artificial seed

Objective:To study the in vitro germination and plantlet regeneration from artificial seeds of Cymbidium aloifolium(C.aloifolium),a highly threatened medicinal orchid of Nepal.Methods:Artificial seeds were produced in vitro by encapsulation of protocorms with 4%sodium alginate and 0.2 mol/L calcium chloride solution.In vitro germination and plantlet regeneration of the artificial seeds were tested by culturing them on different strength of Murashige and Skoog(MS) liquid media(0.25,0.5 and 1.0) and MS liquid medium supplemented with 0.5 mg/L benzyl amino purine and 0.5 mg/L naphthalene acetic acid.Freshly produced artificial seeds were stored up to 28 d at 4 ℃.In order to check the viability,storage artificial seeds were treated with five different sterilization techniques(T_1 T_2,T_3 T_4,T_5) and inoculated on full strength(1.0) of MS liquid medium after each 7 d of interval upto28 th days.Results:The highest percentage of germination(100%) of artificial seed was obtained on quarter(0.25),half(0.5) and full(1.0) strength of MS liquid medium.Experimentally,full strength of MS liquid medium was more effective for earlier seedling development of C.aloifolium.Artificial seeds were successfully stored at 4 ℃ till 28 th days.Treatments T_1 and T_2showed 97.5%viability of storage artificial seeds and hence considered as the most effective sterilization techniques to recover the plant from storage artificial seeds.Plantlets developed from artificial seeds were successfully acclimatized in potting mixture containing cocopeat,litter and sphagnum moss with 85%survival rate.Conclusions:The present study revealed that artificial seeds are the good alternative explants for in vitro mass propagation and short term conservation of C.aloifolium.

Fuzzy Analysis Method for Orthogonal Test on Seed Propagation of Eurya chinensis

Fuzzy analysis method for orthogonal test was used to study the effect of different sowing depth, seed soaking time, illumination intensity and culture soil on the seed germination rate of Eurya chinensis. The results showed that the effect of seed soaking time on the sowing and seedling of Eurya chinensis was the biggest, and the ethers were illumination intensity, sowing depth and culture soil. The germi- nation rate was the highest when composite soil was used as culture soil and seeds were soaked for 48 h and sowed in topsoil, and a layer of sunshade net was used.

Study on Cutting Propagation of Toona ciliata Clones

Cutting propagation rhizogenesis of Toona ciliata clone cutting orchard were studied, and the results were as fol ows： Rooting type included both cal us rooting and cortex rooting. About 17 in 22 of clones were dominated by cortex rooting, accounting for the 77% of clones. There were 5 clones which occurred with both cal us rooting and cortex rooting during rooting process. The rooting efficiency index （REI） showed a positive correlation with the rooting rate. Under natural tem-perature condition, keeping moisture and shading, we found that the highest survival rate occurred in early June, September, and October.

Vegetative propagation of Litsea monopetala, a wild tropical medicinal plant: Effects of indole-3-butyric acid (IBA) on stem cuttings

In this study we investigated the rooting ability and the growth performance of juvenile single-node leafy stem cuttings of Litsea monopetala （Roxb） Pers. collected from two mature mother trees preserved in the hill forest of Chittagong district, Bangladesh. The rooting ability of cuttings was studied under 0%, 0.1%, 0.2% and 0.4% indole-3-butyric acid （IBA） treatments. Significantly better rooting response （p ≤ 0.05） was observed with 0.1% IBA compared to control （0% IBA）. The mean number of roots and the length of the longest root of cuttings in different treatments showed no significant difference （p≤0.05）. After transfer into polythene bags from non-mist propagator, rooted cuttings treated with 0%, 0.1% and 0.2% IBA demonstrated the highest （100 ± 0.00%） survival capacity. The mean number of shoots developed in cuttings in the polythene bags in first three weeks varied significantly （p≤0.05） among the treatments. Effects of three fertilizer treatments, viz. T0 （no fertilizer）, T1 （10g Urea, 20g TSP, 10g MOP dissolved in 1 L water） and T2 （10g Urea, 20g TSP, 10g MOP dissolved in 2 L water） on initial growth ofstecklings were also measured over a 90-days period. The increment of leaf area of stecklings was significantly higher （p≤0.05） under T0 compared with that under T1 and T2 while the increment of stem length, collar diameter and root biomass varied insignificantly among different fertilizer treatments. The results suggest that rooting juvenile single-node leafy stem cuttings could be an effective mean of regenerating L. monopetala. The application of 0.1% IBA concentration is recommended for rooting of juvenile leafy stem cuttings and application of fertilizer appeared unnecessary for the subsequent growth of stecklings in polythene bags.

Correlation of Leaf and Root Senescence During Ripening in Dry Seeded and Transplanted Rice

Dry seeding is a resource-saving rice establishment method. With an equivalent yield, dry seeded flooded rice （DSR） has been considered as a replacement for traditional transplanted flooded rice （TFR）. However, the differences in leaf and root senescence during grain filling between DSR and TFR were seldom identified. In this study, the root length, root tip number and leaf senescence of rice varieties Huanghuazhan and Yangliangyou 6 during ripening were compared between DSR and TFR. Results showed that top three leaves in DSR had the characteristics of relatively lower SPAD value, lower N content and premature leaf senescence. In addition, both the total root length and total root tip number of DSR were significantly lower than those of TFR. In conclusion, premature and quick leaf senescence was related with inadequate root length and root tip number during ripening, which might result from the deficiency of nitrogen supply in DSR. Techniques on improving leaf nitrogen status and delaying the leaf senescence during grain-filling in DSR should be developed in future researches.

In vitro propagation of a medicinal plant: Tripterygium wilfordii Hook f.

In this study a reliable protocol was developed for the establishment of commercial in vitro cultures of Tripterygium wilfordii Hook f.. Juvenile shoots from one-year-old elite plants were used as the source of explants. New axillary shoots were obtained after 30 days of culture on a MS medium supplemented with BAP （2.0 mg.L^-1） and NAA （0.1 mg.L^-1）. The optimal multiplication medium was a modified MS medium supplemented with BAP （1.0 mg.L^-1） and NAA （0.1 mg.L^-1）. This yielded a multiplication rate of 2.4 for each subculture. Slightly more than 92% of shoots rooted when cultured on a modified MS medium containing IBA （0.2 mg.L^-1） and activated charcoal （0.5 mg.L^-1）. Activated charcoal promoted both a strong and a high rooting rate during the rooting phase. Plantlets were transferred to pots for a short acclimatization stage in a greenhouse where 95% of the plantlets survived. This highly reproducible procedure can be adopted for large-scale propagation of T. wilfordii.

Vegetative propagation of Aesculus indica through stem cuttings treated with plant growth regulators

Cuttage propagation of Aesculus indica was tested by treatments with different concentrations of indole-3-acetic acid(IAA) @ 2000, 4000(ppm), indolebutyric acid(IBA) @ 2000, 4000(ppm) and naphthlcetic acid(NAA) @ 2000, 4000(ppm) in dry formulation in the Forest Nursery, Faculty of Forestry, SKUAST-K, Shalimar.The cuttings treated with IBA @ 4000 ppm and IBA @ 2000 ppm had a sprouting rate of 75% and 50%, respectively, which was significantly higher than that of control and other treatments.The highest rooting rate(50%) was recorded in the cuttings with the application of IBA @ 4000 ppm.The cuttings treated with IBA @ 2000 ppm had 25% rooting rate.All other treatments along with control(talc powder) failed to induce rooting.It was concluded that IBA @ 4000 ppm was a better-applied concentration for vegetative propagation of A.indica under Kashmir conditions.

Clonal propagation of Bambusa vulgaris by leafy branch cuttings

Bambusa vulgaris Schrad ex wendl is a widely cultivated bamboo species in rural Bangladesh for its versatile uses. The vegetative propagation becomes the only viable alternative for this species because B. vulgaris does not set seed after sparse flowering, which makes seedling progenies unavailable. A low-cost propagation trial was conducted to explore the clonal propagation techniques for the species with two types of small branch cuttings, nodal leafy cuttings and tip cuttings. The cuttings, after treating with 0, 0.1%, 0.4%, and 0.8% IBA solutions, were kept in non-mist propagator to let them to root for assessing the rooting ability. The cuttings were rooted in four weeks and were allowed to grow in the polybags for 10 months under nursery condition to assess their steckling capacity. The study reveals that both types of branch cuttings are able to develop roots, shoots, to survive and to form rhizome under the nursery condition. Rooting ability of the cuttings was significantly enhanced by the application of rooting hormone - IBA. The highest rooting percentage in nodal leafy cuttings and the tip cuttings （56.67% and 51.0%, respectively） were observed in 0.8% IBA treatment, followed by 0.4% IBA and the lowest （34.3% and 30.0%, respectively） was in control. The highest number of root developed per cutting （9.77 and 8.33 in nodal leafy cuttings and the tip cuttings, respectively） was also obtained from the cuttings treated with 0.8% IBA solution, followed 0.4% IBA treatment and the lowest （3.1 and 2.1, respectively） was in the cuttings without treatment. However, the length of the longest root varied significantly neither with the cutting types nor the concentrations of IBA solution. Survival percentage of the stecklings in nursery condition was significantly enhanced by IBA.

Successful vegetative propagation by stem cuttings of Tetraena mongolica Maxim.

Techniques have been developed for the vegetative propagation of Tetraena mongolica,a rare and endangered species endemic to the western Ordos Plateau in the northcentral portion of China.The purpose of the study was to investigate the rooting characteris-tics of the plant species from the aspects of length and ages of cuttings,external hormone concentration,leaf retention,cutting orientations,and cutting positions.The results obtained from the experiments indicated that the rooting ratio of 2-year or 3-year old cuttings was larger than more than of 4-year old cuttings.The influence of external hormone concentration on rooting was ob-vious;the rooting ratio of cuttings was increased by external hormone treatment,especially when treated by ABT1(ABT No.1 Rooting Powder) 50 mg/L for 3 hours,in which case,the rooting ratio reached 91 percent.Leaf retention influenced rooting sig-nificantly;the rooting rate of cuttings became greater as the leaf retention increased.There was no significant difference in rooting capability between south and north orientations of the ortets.The rooting ratio of the cuttings obtained from the distal portions of shoots was significantly larger than those obtained from the middle and the basal portions of the shoots.On day three after treat-ment,calluses started appearing at the incisions of some cuttings dipped in 500 mg/L IAA(indole acetic acid).Most cuttings were healed or swelled,and 6 percent of cuttings rooted on day 6 after treatment.Higher rooting period occurred 7-10 days after the experiment was set up.

Vegetative propagation of Holarrhena pubescens,a wild tropical medicinal plant:effect of indole-3-butyric acid (IBA) on stem cuttings

The present study investigated the rooting ability and growth performance of juvenile single-node leafy stem cuttings of Holarrhena pubescens, collected from two mature mother trees preserved in the hill forests of Chittagong District, Bangladesh. The rooting ability of cuttings was studied with 0, 0.1%, 0.2% and 0.4% indole-3-butyric acid （IBA） treatments. The better rooting response was observed with 0.1% IBA, but was insignificant compared to other treatments. The mean number of roots and length of the longest root of cuttings in different treatments showed no significant difference. After transfer to the polythene bags from a nonmist propagator, rooted cuttings treated with 0.4% IBA demonstrated the highest （50.00± 0.78%） survival capacity. Regarding initial growth of established rooted cuttings （stecklings） over a 90 d period, the increment of stem length of stecklings under T2 （10 g Urea, 20 g TSP, 10 g MOP dissolved in 2 L water） and leaf area expansion of stecklings under T1 （10 g Urea, 20 g TSP, 10 g MOP dissolved in 1 L water） was significantly higher （p 〈 0.05） compared with that under TO （no fertilizer）. The increment of collar diameter and root biomass production varied insignificantly among different fertilizer concentrations, although stecklings under T2 showed the better performance. The results suggest that rooting juvenile single-node leafy stem cuttings can be an effective mean of regenerating H. pubescens. The application of 0.1% IBA or without IBA is recommended for rooting juvenile leafy stem cuttings. Application of fertilizer appears necessary for subsequent growth of stecklings in polythene bags.

<i>In Vitro</i>Mass Scale Propagation of Wild <i>Cymbidium lowianum</i>with a Rare and Endangered Plant

The wild Cymbidium lowianum, a national-level rare and endangered species of Orchid, is an excellent garden plant with ornamental flowers with striking, deep red lips in a V-shaped formation. The objective of this study was to establish a micropropagation protocol system via immature seeds of wild Cymbidium lowianum, evaluate the Murashige and Skoog (MS), half-strength Murashige and Skoog (1/2 MS) medium, 6-benzylaminopurine (BA), a-napthaleneacetic acid (NAA), organic additions activated charcoal (AC) and banana pulp (BP) effects on the different morphogenesis (seed germination, multiple shoot and rooting) in vitro. The optimal combination for the germination of seed was 1.0 mg·L-1 BA with 0.5 mg·L-1 NAA in 1/2 MS, and addition 0.3%AC, which resulted in 95% seed germination in 90 days. The best formulation for multiple shoot was 1/2 MS medium containing 2.5 mg·L-1 BA, 0.5 mg·L-1 NAA and addition 8% BP in which produced 19.8 shoots per protocorm in 60 days. Multiple shoots were cut and rooted in 1/2 MS supplemented with 1.5 mg·L-1 NAA,0.1 mg·L-1 BA and 0.3% AC, roots initiated 20 days after culture, the rooting percentages reached to 100%, in which 4.7 per shoot produced roots in 60 days. The survival rate of plantlet was up to 92% in moss after 30 days. This finding reveals that it is possible to obtain in vitro culture of Cymbidium lowianum using immature seeds in asymbiotic culture.

Tissue Culture and Gemmae Propagation of Shoot Tipof Huperzia serrata

To solve the problems of microbial contamination and no rooting during the tissue culture of Huperzia serrata, wild seedlings of Huperzia serrata were cultured by indoor hydroponics or soil cultivation methods, then the stem tips were used in tissue culture. The above operation significantly reduced microbial contamination during tissue culture process. Different types and concentrations of hormones were added into the basic medium MS to screen the optimal formula for induced plantlet growth and rooting. It was found that 1/2MSCO was the best medium on which stem, leaves and roots of tissue culture plantlets of Huperzia serrata grew quickly. At the same time, sand planting was a convenient and effective approach for gemmae propagation of Huperzia serrata, the survival rate of seedlings germinated from gemmae was nearly 100%.

Implanting iodine-125 seeds into rat dorsal root ganglion for neuropathic pain: neuronal microdamage without impacting hind limb motion

The use of iodine-125 （L251） in cancer treatment has been shown to relieve patients＇ pain. Consid- ering dorsal root ganglia are critical for neural transmission between the peripheral and central nervous systems, we assumed that 125I could be implanted into rat dorsal root ganglia to provide relief for neuropathic pain. 125I seeds with different radioactivity （0, 14.8, 29.6 MBq） were im- planted separately through L4-5 and L5-6 intervertebral foramen into the vicinity of the L5 dorsal root ganglion, von Frey hair results demonstrated the mechanical pain threshold was elevated after implanting 125I seeds from the high radioactivity group. Transmission electron microscopy revealed that nuclear membrane shrinkage, nucleolar margination, widespread mitochondrial swelling, partial vacuolization, lysosome increase, and partial endoplasmic reticulum dilation were visible at 1,440 hours in the low radioactivity group and at 336 hours in the high radio- activity group. Abundant nuclear membrane shrinkage, partial fuzzy nuclear membrane and endoplasmic reticulum necrosis were observed at 1,440 hours in the high radioactivity group. No significant difference in combined behavioral scores was detected between preoperation and postoperation in the low and high radioactivity groups. These results suggested that the mechan- ical pain threshold was elevated after implanting 125I seeds without influencing motor functions of the hind limb, although cell injury was present.

The Effect of Sample Date and Timing of Cuttings for Maximum Propagation Efficiency of the Grape, <i>Vitis aestivalis</i>“Norton/Cynthiana”

Vitis aestivalis “Norton/Cynthiana” is a grape native to North America and utilized in the commercial wine industry. The purpose of this research was to determine the appropriate time of year and/or time after last freeze that will yield the most successful propagation rate for Vitis aestivalis. It was hypothesized that the highest rate of propagation would be in early summer. During a year-long study (2015-2016), forty cuttings were taken semi-monthly from a vineyard, treated with 0.1% IBA and placed in a plant growth room set to spring conditions. Daily temperatures were recorded every day as well. In a follow-up study, cuttings were taken weekly from early spring before vine budding until late fall (2017) and treated as previous. After six weeks, cuttings were evaluated for root production. In both studies, rooting success rates were the highest in the month of June at 10.0% - 27.5%, with rates less than 7.5% for all other sampling dates. It was determined that the best time to propagate Vitis aestivalis is in June, 10 - 12 weeks after the last temperature below 0°C or eight to ten weeks after the first budding.

Effects of Different Treatments on Rooting and Germination of Paeonia rockii T.Hong et J.J.Li Seeds

[Objectives] This study was conducted to investigate the effects of different treatments on rooting and germination of Paeonia rockii T. Hong et J.J.Li seeds. [Methods] Different stratification time and temperatures and different GA_3 concentrations and treatment time were set to screen the optimum treatment method for promoting seed rooting and germination. [Results] The rooting rate was the highest when the seeds were subjected to 30 d of stratification treatment in an incubator at 25℃, and treated with 300 mg/L GA_3 for 24 h. Soaking the seeds with 300 mg/L GA_3 for 24 h combined with 30 d of stratification treatment was most beneficial to germination, and the germination rate was over 90%. [Conclusions] This study provides technical guidance and basic information for the rooting and germination of P. rockii seeds.