A Preliminary Study on Tissue Culture and Rapid Propagation of Virus-free Plantlets of Benihoppe Strawberry

[ Objective] This study aimed to investigate the tissue culture and rapid propagation techniques of Benihoppe strawberry. [ Method] Shoot tips of new stolons of Benihoppe strawberry were used as experimental materials to analyze the effects of media type, cytokinin type and concentration, carbon source type and concentration, and light intensity on tissue culture and propagation of Benihoppe strawberry. [ Result] MS was the optimal media for plantlet propagation. In the media containing 1.2 ing/L BA （with addition of 0.1 mg/L NAA）, fresh weight, dry weight and propagation coefficient of strawberry plantlets reached the maxi- mum, which were 2.259 g, 0. 221 g and 12.4, respectively. The optimal carbon source was 30 g/L sucrose, and the optimal light intensity was 1 600 lx. [ Conclu- sion] The best media for tissue culture and rapid propagation of Benihoppe strawberry was MS ＋ BA 1.2 mg/L ＋ NAA 0. 1 mg/L ＋ sucrose 30 g/L ＋ agar 8 g/L. This study provided scientific basis for large-scale production of Benihoppe strawberry.

Establishment of Three Breeding System of Virus-free Strawberry Plantlet in Infantry Regiment of the Twelve Division

Shoot-tip culture of strawberry is the most important way to get virus-free plants.Moreover,detoxification and rapid propagation are the key techniques for producing high-quality seeds and seedlings massively and rapidly and achieving the high-quality,high-yield and high-efficiency development of strawberry industry.We have recently produced virus-free strawberry seedlings by combing tissue culture technology and stolon-reproduction method and this technology has been widely used.In addition,a relatively perfect three breeding system of virus-free strawberry seedlings was established after production practices,guaranteeing the supply of high-quality seeds and seedlings and technical support for strawberry industry in corps.

Rapid Propagation of Virus-free Sugarcane Plantlets via Temporary Immersion Bioreactor System

By employing temporary immersion bioreactor system(TIBs),we studied virus-free culture of seedlings from sugarcane varieties ROC16 and ROC22,from medium recipe,inoculation amount,sucrose concentration,and variety difference. The results showed,using this method,that proliferation rate of ROC16 improved by 40 times,per flask generated about 800 plantlets; of ROC22 improved by 30 times,per flask generated about 400-600 plantlets. The results provided basis for using TIBs in rapid propagation of plantlets via tissue culture.

Standardized Production Technology of Virus-free Seed Potato

The research reviewed standardized production technology of virus-free seed potato, including stem tip isolation, tissue culture seedling production, breeder＇s seed and protospecies production, and standard potato tubers＇ production, and concluded a set of standardized production technology for virus-free seed potato, in order to provide theoretical references for improving potato yield.

Evaluation of the Characteristics of Virus-free Sweetpotato and Its Use in Hybridization

Application of virus-free sweetpotato is a breakthrough of sweetpotato production. Several popular varieties were adopted to study their yield increase potential and their use in breeding in this research. The results showed that after virus-elimination, all varieties had a yield increase ranging from 14.95% - 91. 61% and marketable quality improvement, despite the changes in location and season. The changes of dry matter content and disease resistance were not significant. Virus-free sweetpotato performed vigorous vine growth and strong dry matter assimilation ability. Virus-free parents had a slightly high seed setting percentage, but its flowering ability and performance of off-springs were similar to that of its check plants.

Detection of Ratoon Stunting Disease in Virus-free Seedcane via Real-time Fluorescence Quantitative PCR

This study was to develop the real-time fluorescence quantitative PCR technique for detecting the ratoon stunting disease （RSD） in virus-free seedcane seedlings. Healthy tissue culture seedlings were obtained from six plants of sugarcane ROC22, which had been confirmed RSD-positive by detecting the sugarcane juice, by employing the sugarcane seedlings production protocol. Real-time fluorescence quantitative PCR was used to detect RSD pathogens in tissue culture sam- pies. The results showed that target fragment of RSD pathogens was not found in all 10 samples in real-time fluorescence quantitative PCR, with the Ct values of 37 - 39. The healthy tissue culture sugarcane seedlings do not carry RSD pathogens, indicating that adopting healthy seedcane seedlings production technique could thoroughly get rid of RSD pathogens.

Effect of Heat Treatment Combined with Shoot Tip Culture on the Virus-Free of Arena Strawberry

With Arena strawberry as experimental materials, heat treatments combined with shoot tip culture were used to explore their detoxification effects. The results indicated that if not heated, splitting shoot tips directly could not completely remove viruses. On the other hand, it will be obviously improved after shoot tips are heated. Incubators heat treatment （inconstant temperature） with shoot tip culture, constant temperature water bath incubation process with meristem-tip culture, hot air treatment （constant temperature） with apical meristem culture were three methods tested in searching for virus-free of Arena strawberry in this experiment. And their virus-free rates were 78.9%, 86.0% and 50.0% respectively. Of the three methods, the second, constant temperature water bath process with shoot tip culture, with 78.7% survival, is the best. Thus, it can be inferred that constant temperature water bath treatment combined with shoot tip culture might be the most appropriate method to remove viruses of Arena strawberry.

Expression of Invertase Genes in Virus-free Sugarcane

Virus-free sugarcane seedlings have improved biomass and sucrose content compared with ordinary seedlings, and sucrose invertases are key enzymes regulating sugarcane growth and sucrose accumulation. In this study, the differences in the expression levels of 3 invertase genes, CWI, SAI and NI, between virus- free and ordinary sugarcane seedlings were analyzed. Compared with ordinary sugarcane plants, the expression of CWI was mainly up-regulated in immature leaves and stems at elongation stage and leaves and immature internodes at maturation stage, and especially, greatly up-regulated in immature interuedes at maturation stage of virus-free plants. The expression of SAI and NI were mainly up-regnlated in leaves and immature internedes of virus-free plants at maturation stage, which might be beneficial to sugar accumulation and rapid utilization of monosaccharide in the stalks of virus-free plants. It is further indicated that virus-free treatment could significantly improve the expression of sucrose invertases at late growth period, and might facilitate the increase of plant biomass.

Detection of Ratoon Stunting Disease in Virus-free Seedcane of Saccharum officinarum by PCR

[Objective]The study was carried out for providing good method to detect ratoon stunting disease（RSD）in virus-free seedcane of Saccharum officinarum.[Method]PCR method was used for the detection of RSD in virus-free seedcane of sugarcane.[Result]During the different growth stages of healthy virus-free seedling including proliferation stage,rooting stage,sand culture stage and temporary planting stage,the RSD detection results were negative.PCR detection sensitivity in the present study showed that PCR could detect RSD pathogen in the solution with bacterium concentration 10-3.[Conclusion]PCR method showed a good specificity and higher sensitivity,so it was suitable for the batch detection in the production of healthy virus-free seedlings of sugarcane.

Study on Virus-free Breeding Technology of Potato Microtuber

It is the simplest,fastest and effective way to improve the yield level of potato by selecting high quality virus-free seed potato for field production and maintaining the original characters of virus-free minituber. Microtuber is featured by small volume,no pathogen,convenient storage and transportation,which can be induced in summer with high temperature and humidity,and overcome a series of problems such as poor survival rate of transplanted test-tube plantlets,and easy pollution of cutting tips. Therefore,it can replace conventional test-tube plantlets directly for reproduction. Meantime,if minitubers are sown ahead of time using cold frame,the growth stage of potato minituber will avoid the occurrence period of aphids. The use of gauze cover for insect prevention has good effect on maintaining the excellent characters of microtuber. The yield and quality of virus-free potato are greatly improved by means of virus-free culture of shoot tip seedlings,subculture propagation of virus-free seedlings,induction of microtubers by dark culture,cultivation of breeder’s seeds in greenhouse and screenhouse to preserve the characters of potato.

Correlations between delayed fluorescence of chlorophyll, metabolism and yield of plants. III. Influence of viral infection on field plants and new technology of clone selection of virus-free planting potato

At the end of potato plants vegetation virus in-fection induced both decrease in maximum amplitude of delayed fluorescence maximal amplitude and increase half time of its decrease, as well as reduction in the amount of stems, plants’ height and assimilation area surface, yield, acceleration of plants development and their early die-off. The differences of DF pa-rameters and yields between strongly and weakly infected plants increase in case of a combined virus infection. In industrial test of the selection of virus-free planting potato by the use of DF parameter, a rise in the yield and de-crease degree of viral infection of crops was obtained.

Growth and Transcriptomics Analysis ofMichelia macclurei Dandy Plantlets with Different LED Quality Treatments

Michelia macclurei Dandy is a significant tree species that has extensive cultivation for forestry and horticulture purposes in southern China,owing to its economic and practical importance.Light quality influences plantlet growth and development during tissue culture.However,the growth characteristic and molecular regulation of M.macclurei under different light quality conditions are not well understood yet.In this study,we investigated the morphological,chlorophyll content,and transcriptomic responses of M.macclurei plantlets under different light-emitting diode(LED)qualities,including white,blue,and red light.The results showed that blue light significantly increased plant height(21.29%)and leaf number(18.65%),while red light decreased plant height and leaf number by 7.53%and 16.49%,respectively.In addition,the plantlets’chlorophyll content and etiolation rate were significantly reduced by blue and red light quality compared to white light.Compared to white light,blue light had a negative effect,leading to decreased rooting rate(64.28%),root number(72.72%),and root length(75.86%).Conversely,red light had a positive effect,resulting in increased rooting rate(24.99%),root number(109.58%),and root length(72.72%).Transcriptome analysis identified 54 differentially expressed genes(DEGs)in three groups that consisted of blue light vs.white light(BL-vs-WL),red light vs.white light(RL-vs-WL),and red light vs.blue light(RL-vs-BL).Specifically,21,7,and 41 DEGs were identified in the three groups,respectively.The DEGs found in the RL-vs-WL and BL-vs-WL groups were involved in plant hormone signaling,nitrogen metabolism,and phenylpropanoid biosynthesis pathways,which suggests that M.macclurei plantlets adapt to the changes of light quality via modulating gene expression.Overall,our study provides valuable insights for understanding the molecular and morphological responses of M.macclurei plantlets under different light qualities.

Evaluation of Different Substrates Compositions for Acclimatization of Tissue Culture Taro Plantlets in a Propagator

Taro is cultivated in most Regions of Cameroon and it is affected by taro leaf blight disease since 2010 which has decreased its production. Lack of disease-free planting materials has been a main problem to farmers. This study was carried out at International Institute of Tropical Agriculture (IITA) Yaounde and Institute of Agricultural Research for Development (IRAD) Bambui to assess different substrates for acclimatization of tissue culture taro plantlets in apropagator. No information is available on acclimatization of Cameroonian taro plantlets in different substrates. Taro plantlets from tissue culture were acclimatised in a propagator for six weeks under different substrates, the first substrate consisted of sterile three parts of soil and one part of river sand mixed together (3:1), the second substrate consisted of sterile two parts of soil and two parts of river sand mixed together (2:2), the third substrate consisted of sterile two parts of soil, one part of rice husk and one part of river sand mixed together (2:1:1) and the fourth substrate consisted of sterile one part of soil and three parts of river sand mixed together (1:3). After acclimatisation of the different taroplantlets (Dark green petiole with small leaves (L1), Red petiole with small leaves (L2), Light green petiole with large leaves (L3) and Light green petiole with small leaves (L4) in these four substrates, it was observed that the best growth rate of plant was recorded on substrate sand + soil (1:3). The other substrates showed moderate growth of plants. Substrate sand + soil (1:3) can be recommended for acclimatization of Cameroonian taro plantlets.

Steps To ward HIV Cure Two more HIV-infected patients appear to be virus-free after bone-marrow transplants

THIS year＇s International AIDS Society Conference on HIV Pathogenesis, Treatment and Preven-／tion, held in Kuala Lumpur, Malaysia, made major headlines when Timothy Hendch, an American doctor, announced that two more cancer patients may have been cured of HIV after receiving bone-marrow transplants to treat lymphoma. Both patients had been taking retroviral medication, and continued to do so after the transplants as their viral levels sank until doctors were unable to find any traces of HIV in the patients＇ blood.

橡胶树体胚苗质量调查与分析

天然橡胶是我国重要的战略物资和工业原料,主要来源于橡胶树。橡胶树生长周期长,其苗木质量直接影响天然橡胶产业的发展,橡胶树体胚苗在前期研究中表现一致性好,且具有速生、高产的优势。海南省天然橡胶新型种植材料创新基地是当前规模生产的橡胶树体胚苗的基地,现生产品种有热研73397和热研917。本研究分析比较了热研73397体胚试管苗在2018—2020年间的株高、茎粗、叶片数、主根长等指标的差异性,比较了2020年2个品种各指标的差异性,并对不同年份不同品种进行分级比较,以期阐明橡胶树体胚苗规模生产的情况。结果表明:(1)2018—2020年,热研73397的株高、茎粗、叶片数、主根长度在各区间的比例相当;株高、叶片数、主根长度在3 a间的差异不显著;茎粗在2018和2019年之间差异显著,且3 a均值均在2.00 mm以上,最大值和最小值的差值仅为0.09 mm。(2)2020年,热研73397和热研917体胚苗的株高一致,均值均为8.0 cm,且大部分集中在5.0~10.0 cm,分别占66.13%、66.91%;热研73397和热研917的茎粗范围分别为0.80~4.23 mm、1.13~3.73 mm,均值分别为2.11 mm和2.18 mm,2个品种的茎粗在1.50~3.00 mm之间高度集中,分别占85.94%、83.82%;热研73397和热研917叶片数范围分别为0~6片和1~4片,2个品种叶片数绝大多数为1~3片,分别占88.82%、96.32%,其中热研917叶片数为2片的占比高达52.94%;热研73397的主根长度均值为9.5 cm,而热研917的为9.1 cm,2个品种的主根长度多数集中在8.0~12.0 cm之间,分别占74.12%、79.41%;方差分析结果表明,2020年2个品种的株高、茎粗、叶片数差异不显著,主根长度差异显著(P<0.05)。(3)2018—2020年调查的橡胶树体胚苗中,不同年份不同品种的橡胶树体胚苗,一级苗比例为19%~22%,二级苗比例为56%~62%。据此,规模化生产橡胶树体胚试管苗质量稳定,为橡胶树体胚苗生产工艺的推广应用提供参考。

γ-氨基丁酸对怀玉山三叶青低温胁迫下试管苗生长及相关生理代谢的影响

目的探讨γ-氨基丁酸(GABA)对怀玉山三叶青低温胁迫下试管苗生长及相关生理代谢的影响,为其在怀玉山三叶青仿野生栽培中的防冻抗寒应用提供依据。方法以低温胁迫下GABA处理的怀玉山三叶青试管苗为研究对象,采用植物组织培养法测定其生长指标,植物生理生化法测定其生理指标,实时荧光定量聚合酶链反应(qRT-PCR)法检测其查尔酮合酶和黄酮醇合酶基因的相对表达量。结果与未添加GABA的对照组相比,培养基中添加0.5 g·L^(-1)GABA,怀玉山三叶青2个栽培种“怀玉1号”和“怀玉2号”试管苗的增殖系数、株高、鲜质量、干质量及组织含水量均达到最高值;可以显著提高其可溶性蛋白(SP)、可溶性糖(SS)、脯氨酸(Pro)、还原型谷胱甘肽(GSH)、抗坏血酸(AsA)、吲哚乙酸(IAA)、玉米素核苷(ZR)、赤霉素(GA3)、茉莉酸(JA)的含量,以及超氧化物歧化酶(SOD)、过氧化物酶(POD)、抗坏血酸过氧化物酶(APX)、谷胱甘肽还原酶(GR)的活性和查尔酮合酶(CHS)基因的相对表达量(RQ值);显著降低其相对电导率、脱落酸(ABA)、过氧化氢(H_(2)O_(2))、丙二醛(MDA)的含量,O_(2)·^(-)产生速率,以及黄酮醇合酶(FLS)基因的相对表达量。低温胁迫下GABA处理的怀玉山三叶青试管苗的生长与IAA、ZR、GA3、JA、SS、AsA、GSH的含量和CHS基因表达呈正相关。结论适当浓度的GABA可以调控内源植物激素和渗透调节物质含量、抗氧化物质含量以及次生代谢物关键基因表达,以促进低温胁迫下怀玉山三叶青试管苗的生长。

Effects of Sodium Chloride Stress on Growth of Sweet Potato Plantlets in vitro and Ion Content

[Objective] This study aimed to investigate the salt-tolerance mechanism of sweet potato.[Method]Two sweet potato varieties of Xu 25-2(salt-tolerant cultivar)and Triumph 100(salt-sensitive cultivar)were treated by sodium chloride with the concentration of 0 mmol/L and 100 mmol/L.After 20 days,Na+ content and Na+/K+ ratio in the roots,shoots and leave were determined by the flame photometer,while dry weight and fresh weight of roots,shoots and leave in different varieties were also studied.[Result]The growth of two sweet potato varieties was inhibited under salt stress,so the plant became shorter,leaf and root became fewer,dry weight of roots and leave decreased,but seedlings of Xu 25-2 were inhibited slightly.Furthermore,Na+ content and Na+/K+ ratio in roots,shoots and leaves of two sweet potato varieties increased.Na+ content of salt-tolerant Xu 25-2 was low in roots,shoots and leaves,while Na+ content of salt-sensitive Triumph 100 was high in shoots and leave of seedlings,but the change range of Xu 25-2 was less than that of Triumph 100.[Conclusion]The lower Na+ content and Na+/K+ ratio in leaves under salt stress were the most important characteristics for salt-tolerance of sweet potato varieties.

Callus induction from leaves of different paulownia species and its plantlet regeneration

The experiment was carried out on five different species of Paulownia for callus induction from leaves. MS medium was adopted as basic medium, and from different combinations of NAA and BA the suitable media were determined for callus induction, bud differentiation, and root differentiation of five different species. MS+0.5NAA+4BA, MS+0.3NAA+2BA, MS+0.5NAA+4BA, MS+0.3NAA+6BA, and MS+0.3NAA+8BA were suitable media of callus inductions of leaves, respectively, for Paulownia tomentosa, Paulownia australis, Paulownia fortunei, Paulownia elongata and P. tmentosa x P. fortunei, and MS+0.3NAA+12BA, MS+0.3NAA+12BA, MS+0.5NAA+12BA, MS+0.5NAA+12BA, and MS+0.7NAA+12BA were suitable media for bud differentiation from leaf callus respectively for above five species. The rooting media was determined as 2MS+0.1NAA, 1/2MS+0.1NAA, 1/2MS, 1/2MS+0.3NAA, and 1/2MS+0.5NAA. These results provide reference data for breeding new fine va-rieties with different kinds of Paulownia protoplasts fusions.

碳离子束辐照对马铃薯组培苗生长及生理特性的影响

为研究高能碳离子束对马铃薯组培苗茎切段的影响,以马铃薯品种‘新大坪’的组培苗茎切段为研究材料,设置4个辐照剂量梯度对其进行辐照处理,测定植株致死率、生根率、侧芽形成率、形态变异率以及叶片生理指标,确定‘新大坪’组培苗茎切段的半致死辐照剂量。结果表明:辐照剂量与致死率之间的关系呈正相关性;组培苗茎切段的再生植株表型变化主要为叶片不对称、叶缘不规则、茎变异伸长、叶片丛生和叶片粘连等;随辐照剂量升高,超氧化物歧化酶、过氧化物酶和过氧化氢酶活性呈先升后降的趋势;丙二醛含量则呈先升后降,随后又升高的现象。利用线性回归方程计算‘新大坪’的半致死辐照剂量为20.37 Gy。

珙桐体细胞胚胎发生技术研究

研究探讨基本培养基类型对珙桐愈伤组织诱导的影响、植物生长调节剂对胚性愈伤组织诱导及体胚发育的影响,初步建立以未成熟合子胚为外植体,通过体细胞胚胎发生途径再生珙桐植株的培养方案。结果表明,在含有3%蔗糖、0.3%凝胶、800 mg/L水解酪蛋白、400 mg/L L-谷氨酰胺的MS、1/2MS和WPM基本培养基上均可诱导出愈伤组织,以MS+2.0 mg/L 2,4-D+0.5 mg/L 6-BA诱导率最高,可达94.0%;胚性愈伤组织在MS+0.5 mg/L NAA+1.0 mg/L 6-BA的培养基上诱导效果较好;在MS+0.5 mg/L NAA+1.0 mg/L 6-BA培养基上,体胚诱导率可达22.9%;成熟体胚在1/2MS+0.5 mg/L 6-BA+0.25 mg/L IBA的萌发培养基上能正常萌发,其后在添加1 g/L活性炭的萌发培养基上转化成完整小植株。组织细胞学观察表明,珙桐体胚起源于胚性愈伤组织,经历了球形期、心形期、鱼雷期和子叶期发育阶段,这与自然条件下的合子胚形成过程相似。