Evaluating the efficacy of an attenuated Streptococcus equi ssp. zooepidemicus vaccine produced by multi-gene deletion in pathogenicity island SeseCisland_4

Streptococcus equi ssp. zooepidemicus(SEZ) is a pathogen associated with a wild range of animal species. Frequent outbreaks have occurred in recent years in pigs, horses, goats and dogs which is liable to infect humans. There is a lack of efficient vaccines against this disease and the occurrence of antibiotic resistance may render drug therapies ineffective. In this study, gene deletion mutant(ΔSEZ) in pathogenicity islands SeseCisland_4 was constructed. The mutant ΔSEZ had a 52-fold decrease in 50% lethal dose(LD_(50)) and had less capacity to adhere epithelial cells. Importantly, immunization of mice with attenuated vaccine ΔSEZ at the dose of 10~2 colony-forming units(CFU) mL~(–1) elicited a significant humoral antibody response, with an antibody titer of 1:12 800. Therefore, 10~2 CFU mL~(–1) might be used as the appropriate immune dose for the attenuated vaccine ΔSEZ, which provided mice with efficient protection against virulent SEZ. In addition, the hyperimmune sera against 10~2 CFU m L–1 attenuated vaccine ΔSEZ could confer significant protection against virulent SEZ infection in the passive immunization experiment and exhibited efficient bactericidal activity in the whole blood assay. Meanwhile, no viable bacteria was detected in blood when mice were immunized with ΔSEZ at the dose of 10~2 CFU mL~(–1) via hypodermic injection. Thereafter, the mutant ΔSEZ at the dose of 10~2 CFU mL~(–1) could confer significant protection in mice and had less negative effects on host, which could be an effective attenuated vaccine candidate for the prevention of SEZ.     

两种LD50计算方法对副溶血性弧菌毒力的比较研究

副溶血性弧菌(Vibrio parahaemolyticus,Vp)是目前备受关注的人畜共患病的病原。为了比较7株不同来源的Vp对斑马鱼的毒力大小及致病能力,采用急性毒性试验测定96 h内Vp对斑马鱼的半数致死剂量(median lethal dosage,LD_(50)),用累积法和Bliss法对比分析LD_(50)指标。结果显示:Vp KNH1无致病性。其他菌株感染后,均表现出腹部出血,腹腔有腹水,内脏出血等症状。用累积法和Bliss法分析人类致病菌株ATCC17802、ATCC33847及水生动物致病菌株Vp13、Vp31、Vp41、Vp57共6株不同来源的副溶血性弧菌的LD_(50),累积法计算得到的LD_(50)分别为6.0×107、7.4×107、3.9×107、1.6×108、1.6×108、9.3×107CFU/m L;Bliss法得到的LD_(50)分别为:5.0×107、7.6×107、3.6×107、1.5×108、1.6×108、9.4×107CFU/m L。研究结果发现两种计算方法得出的LD_(50)数值差距较小,结果相似,不同菌株毒力强弱的顺序均为:Vp13ATCC17802ATCC33847Vp57Vp31≥Vp41Vp KNH1。综上所述,累积法及Bliss法均可用于检测细菌毒力大小,但由于计算机的发展使得Bliss计算方法更加简便,为更优的选择。     

草鱼暴发性出血病病原分离、毒力基因检测与药敏分析

从草鱼体内分离到致病菌株Jxsks1,该菌株对草鱼的LD50值为2.3×103CFU/g鱼体质量,为强致病菌;经生理生化、16S r DNA和gyr B基因序列测序鉴定为嗜水气单胞菌。ERIC-PCR分型结果表明,该菌株为迄今未见报道的类型。选取丝氨酸蛋白酶ahp、热稳定性肠毒素ast、气溶素aer A、热不稳定性肠毒素alt、鞭毛基因fla、脂酶lip等6种毒力基因特异性引物对该菌株进行PCR检测,结果表明其毒力基因型为aer A~+、alt~+、ast~+、ahp~+、lip~+、fla~-。选取28种抗生素进行药敏检测结果表明,该菌株对喹诺酮类、氨基糖苷类、四环素类、氯霉素、第二代和第三代头孢类等药物敏感,对硝基呋喃类、大环内酯类、多肽类中度敏感,对第一代头孢类、青霉素类、复方新诺明和洁霉素耐药。     

Chemical mutagenesis and soybean mutants potential for identification of novel genes conferring resistance to soybean cyst nematode

The resistance of soybean (Glycine max (L.) Merr.) to soybean cyst nematode (SCN, Heterodera glycines Ichinohe), which is a devastating pathogen in soybean production and causes a large quantity of annual yield loss worldwide, can shift during the long-term interaction and domestication. It is vital to identify more new resistance genetic sources for identification of novel genes underlying resistance to SCN for management of this pathogen. In the present study, first, two ethane methylsulfonate-mutagenesis soybean M2 populations of PI 437654, which shows a broad resistance to almost all of SCN races, and Zhonghuang 13, which is a soybean cultivar in China conferring strong resistance to lodging, were developed. Many types of morphological phenotypes such as four- and five-leaflet leaves were observed from these two soybean M2 populations. Second, 13 mutants were identified and confirmed to exhibit alteration of resistance to SCN race 4 through the forward genetic screening of 400 mutants of the PI 437654 M2 population, the rate of mutants with alteration of SCN-infection phenotype is 3.25%. Third, these identified mutants were further verified not to show any changes in the genomic sequences of the three known SCN-resistant genes, GmSHMT08, GmSNAP18 and GmSANP11, compared to the wild-type soybean; and all of them were still resistant to SCN race 3 similar to the wild-type soybean. Taken together, we can conclude that the 13 mutants identified in the present study carry the mutations of the new gene(s) which contribute(s) to the resistance to SCN race 4 in PI 437654 and can be potentially used as the genetic soybean sources to further identify the novel SCN-resistant gene(s).     

Increased ammonification,nitrogenase, soil respiration and microbial biomass N in the rhizosphere of rice plants inoculated with rhizobacteria

Azospirillum brasilense and Pseudomonas fluorescens are well-known plant growth promoting rhizobacteria. However, the effects of A. brasilense and P. fluorescens on the N cycles in the paddy field and rice plant growth are little known. This study investigated whether and how A. brasilense and P. fluorescens contribute to the N transformations and N supply capacities in the rhizosphere, and clarified the effects of A. brasilense and P. fluorescens on the N application rate in rice cultivation. Inoculations with A. brasilense and P. fluorescens coupled with N application rate trials were conducted in the paddy field in 2016 and 2017. The inoculations of rice seedlings included four treatments: sterile saline solution(M_0), A. brasilense(Mb), P. fluorescens(Mp), and co-inoculation with a mixture of A. brasilense and P. fluorescens(Mbp). The N application rate included four levels: 0 kg N ha~(–1)(N_0), 90 kg N ha~(–1)(N_(90)), 180 kg N ha~(–1)(N_(180)), and 270 kg N ha~(–1)(N_(270)). The results indicated that the Mbp and Mp treatments significantly enhanced the ammonification activities in the rhizosphere compared with the M_0 treatment, especially for higher N applications, while the Mbp and Mb treatments greatly enhanced the nitrogenase activities in the rhizosphere compared with the M_0 treatments, especially for lower N applications. Azospirillum brasilense and P. fluorescens did not participate in the nitrification processes or the denitrification processes in the soil. The soil respiration rate and microbial biomass N were greatly affected by the interactions between the rhizobacteria inoculations and the N fertilizer applications. In the Mbp treatment, N supply capacities and rice grain yields showed no significant differences among the N_(90), N_(180), and N_(270) applications. The N application rate in the study region can be reduced to 90 kg N ha~(–1) for rice seedlings co-inoculated with a mixture of A. brasilense and P. fluorescens.     

生物炭对土壤有机质结合铜库容量大小及组成的调控

为研究生物炭对土壤中有机质组分结合铜库容量的大小与组成的调控能力,选择海南省具有代表性的热带次生林和橡胶林的土壤进行模拟实验,根据其有机质含量低、中、高,分别记为SoilL、SoilM、SoilH,以SoilH的含碳量为标准,向SoilL和SoilM中加入玉米秸秆生物炭(处理土壤分别记为SoilL+B和SoilM+B),并通过添加不同浓度的硝酸铜模拟土壤不同铜污染水平。结果表明:SoilL+B和SoilM+B的有机质结合态铜饱和容量分别为8.70 mg·g-1和9.76 mg·g-1,分别为SoilL和SoilM的3.69倍和3.12倍;SoilL+B较SoilL的富啡酸结合态铜(FA-Cu)、胡敏酸结合态铜(HA-Cu)、球囊霉素结合态铜(GRSP-Cu)和颗粒有机碳结合态铜(POM-Cu)的饱和容量分别增加了1.71、2.83、2.75 mg·g-1和0.09 mg·g-1,而黑炭结合态铜(BC-Cu)的饱和容量降低了0.76 mg·g-1;SoilM+B较SoilM的FA-Cu、HA-Cu、GRSP-Cu和POM-Cu的饱和容量分别增加了1.66、3.75、2.68 mg·g-1和0.07 mg·g-1,但BC-Cu的饱和容量降低了2.14 mg·g-1;生物炭对SoilL和SoilM中5种有机组分结合态铜库贡献的大小顺序均为HA>GRSP> FA>POM>BC。研究表明,添加生物炭可以显著提高土壤有机质结合铜库和各有机组分铜库的容量,并提高土壤对重金属的缓冲能力。     

斑马鱼幼鱼嗜中性粒细胞对副溶血弧菌清除的动态变化

副溶血弧菌(Vibrio parahaemolyticus)是人-兽-鱼共患的致病菌,严重威胁食品安全和人类的健康。为了研究宿主嗜中性粒细胞清除细菌感染的动态变化过程,利用红色荧光蛋白标记的副溶血弧菌Vp57~(RFP)菌株感染受精后48 h(hours post fertilization,hpf)的斑马鱼(Danio rerio)幼鱼耳泡,建立局部感染模型,并以大肠杆菌(Escherichia coli)Ec01菌株为对照,系统比较了副溶血弧菌和大肠杆菌的半数致死剂量(median lethal dose,LD_(50))、存活曲线以及和嗜中性粒细胞相互作用的动态过程,RT-qPCR验证炎症相关基因il1b和il10在感染前后的表达量变化。结果显示副溶血弧菌和大肠杆菌感染斑马鱼幼鱼的LD_(50)分别为7.90×10~8 CFU/mL和1.14×10~(11) CFU/mL,与感染大肠杆菌相比,斑马鱼感染副溶血弧菌后招募嗜中性粒细胞的速度更快,且数量更多。RT-qPCR结果发现斑马鱼感染副溶血弧菌后在相同时间点能够激活更高的il1b和il10基因的表达,引起强烈的炎症反应。基于以上研究,我们建立的副溶血弧菌-斑马鱼幼鱼耳泡局部感染模型,为进一步研究嗜中性粒细胞清除副溶血弧菌感染的动态过程,以及深入揭示天然免疫应答机制提供了依据。     

半海水筛选耐盐紫花苜蓿细胞系的生理特性分析

以紫花苜蓿无菌苗下胚轴为外植体,诱导获得愈伤组织(M0)。经甲基磺酸乙酯(EMS)处理,通过一步筛选获得了耐半海水w(NaCl≈1.5%,1/2海水)的耐盐细胞系(Ms1),一部分耐盐细胞系转移到无盐培养基上,继代培养10代后得到新细胞系(Ms2)。无论是在无盐还是不同程度盐胁迫条件下,Ms1的成活率、生长率、脯氨酸含量、Cl-、K 、K /Na 、可溶性蛋白质含量都高于M0,差异显著。Ms2介于两者之间,但是与Ms1基本接近,差异不显著。在w(NaCl≈0.6%,1/5海水)条件,M0细胞系的SOD活性高于Ms1和Ms2;在w(NaCl≈1%,1/3海水)条件下,Ms1、Ms2细胞系的SOD活性达到最高,与M0可溶性蛋白质SDS-PAGE图谱相比较,Ms1、Ms2中出现两条新多肽带(P1=58kD,P2=26kD),这两条多肽带可能与耐盐性有关。     

Inhibition of Na＋,K＋-ATPase in Housefly （Musca domestica L.） by Terpinen- 4-ol and Its Ester Derivatives

To reveal the insecticidal mechanism of terpinen-4-ol, the activity of Na＋,K＋-ATPase in insects tested were determined in vivo and in vitro. The results showed that terpinen-4-ol and its ester derivatives had strong contact activity to housefly and the contact toxicities of its derivatives except Z3 were all superior or equivalent to terpinen-4-ol. All the 7 compounds had strong inhibition towards activity of Na＋,K＋-ATPase. With poisoning symptom exacerbating, the inhibition rates were gradually increased. In vitro, the IC50 of terpinen-4-ol, Z1, Z2, Z4, Z5, and Z6 was 155.89, 197.98, 96.02, 121.36, 124.85, and 153.74 μg mL% respectively. There was well correlation between the LDs0 of terpinen-4-ol derivatives to housefly and the IC50 of terpinen-4-ol derivatives to Na＋,K＋-ATPase in housefly. In conclusion, Na＋,K＋-ATPase was likely the target of terpinen-4-ol against insects.     

Construction and Virulence of Filamentous Hemagglutinin Protein B1 Mutant of Pasteurella multocida in Chickens

Pasteurella multocida, a Gram-negative nonmotile coccobacillus, is the causative agent of fowl cholera, bovine hemorrhagic septicemia, enzoonotic pneumonia and swine atropic rhinitis. Two filamentous hemagglutinin genes, fhaB1 and JhaB2, are the potential virulence factors. In this study, an inactivationfhaB1 mutant ofP. multocida in avian strain C48-102 was constructed by a kanamycin-resistance cassette. The virulence of thefhaB1 mutant and the wild type strain was assessed in chickens by intranasal and intramuscular challenge. The inactivation offhaB1 resulted in a high degree of attenuation when the chickens were challenged intranasally and a lesser degree when challenged intramuscularly. ThefhaB1 mutant and the wild type strain were investigated their sensitivity to the antibody-dependent classical complement-mediated killing pathway in 90% convalescent chicken serum. ThefhaB1 mutant was serum sensitive as the viability has reduced between untreated serum and heat inactivated chicken serum （P〈0.007）. These results confirmed that FhaB1 played the critical roles in the bacterial pathogenesis and further studies were needed to investigate the mechanism which caused reduced virulence of the fhaB1 mutant.     

分子连锁分析探讨家蚕高抗BmNPV品系的抗性遗传基础

【目的】家蚕核型多角体病毒(Bombyx mori nucleopolyhedrovirus,BmNPV)所引发的血液型脓病是一种传染性强的蚕病,极大地影响蚕业生产。本研究旨在定位对BmNPV高抗的家蚕品系中控制其抗性的基因,进而解析其抗性遗传机制,为培育抗性素材和品系提供理论支持。【方法】以BmNPV高抗家蚕品系99R和较感品系Dazao-N为亲本,配制连锁分析(BC_1F)和定位分析(BC_1M)回交群体。首先对两个亲本品系进行浓度梯度添毒,统计感病死亡的家蚕头数,运用SPSS 17.0软件,计算其半致死剂量(LD_(50)),在此基础上确定BC_1分离群体的攻毒剂量,并通过单头定量攻毒,选取感病个体作为连锁定位分析材料;利用筛选得到的覆盖家蚕全套常染色体的多态性标记进行分型,并通过T检验计算各标记与抗性的连锁显著性水平(P值),筛选出与抗性相连锁的标记。在这些标记所在的染色体上加密标记,检测各标记在定位分析群体中的基因型,定位抗性基因。【结果】LD_(50)(99R)=2.92×10~6个多角体/头,LD_(50)(Dazao-N)=9.78×10~5个多角体/头,基于双亲的半致死剂量,选择介于两者之间且略高于其均值的剂量——2×10~6个多角体/头作为BC_1分离群体的攻毒剂量;先后于2014年秋季和2015年春季处理并检测连锁分析群体,前后两次所进行的连锁分析结果有较大差异,其中第一次找到Chr22上的多态性标记S2205与99R抗性连锁,而第二次的连锁分析显示标记S2205与抗性不连锁,也没有找到其他的连锁关系。通过与前人对BmNPV抗性的连锁定位分析结果进行对比,发现连锁定位分析结果的不可重复性是一个普遍的问题。AY380833是GenBank中已公布的在家蚕高抗品系NB和871C中与其抗性位点紧密连锁的分子标记,本研究调查发现其与99R和871C的抗性位点均不连锁。【结论】分子连锁分析结果证明,家蚕对BmNPV的抗性在不同抗性品系中遗传基础有很大的差异性,同一品系可能具有多个抗性位点;家蚕对BmNPV的抗性是一种复杂性状,在符合"质量-数量性状"结论的同时,其数量性状特征突出。     

The Twin-Arginine Translocation （Tat） Pathway Is Essential for Virulence,Flagellation, and Chemotaxis in Xanthomonas oryzae pv. oryzicola Strain RsGD42

Bacterial leaf streak, caused by Xanthomonas oryzae pv. oryzicola, is an important disease of rice （Oryza sativa）. Genetic determinants （tatABC genes） of the twin-arginine translocation （Tat） pathway from X. oryzae pv. oryzicola strain RsGD42 were cloned and characterized, meanwhile, a tatC disruption mutant was generated. The tatC mutant lacked detectable flagella and was highly impaired in motility and chemotaxis. Furthermore, it was observed that the tatC mutant exhibited a reduced production of extracellular polysaccharide （EPS） and a significant reduction of virulence on adult rice plants compared to wild type strain. However, the tatC mutation in X. oryzae pv. oryzieola strain RsGD42 did not affect the growth rate and the ability to induce hypersensitive response （HR） in nonhost tobacco （Nicotiana tabacum L. cv. Samsun）. In conclusion, the data indicated that the Tat pathway significantly contributed to the virulence of X. oryzae pv. oryzicola.     

唐菖蒲复色花突变体AFLP及SCAR标记研究

通过AFLP分子标记方法,应用16个引物组合对唐菖蒲M3突变体和对照株进行基因组序列多态性检测,发现有4对引物扩增出稳定的重复性好的多态性条带,4条特异带暂命名为E4M7304、E6M3202、E6M8258和E8M5268.其中E6M8258和E8M5268为对照缺失条带,其他2条为M3缺失条带.将这些特异性条带回收、克隆、测序,设计SCAR引物进行单株验证,其中E4M7304标记已经转换成了稳定的SCAR标记.由于唐菖蒲从子球到开花种球的繁殖年限较长,故可利用该SCAR标记对唐菖蒲的花色变异突变体后代进行早期分子鉴定,以提高育种效率.     

新疆地区牛分枝杆菌29个毒力基因的突变分析

为了解我国新疆地区牛分枝杆菌的毒力基因变异情况并初步评估其毒力变化,利用聚合酶链式反应(PCR)检测临床分离的66株牛分枝杆菌的29个毒力基因,经Sanger测序后与标准株AF2122/97和本实验室保存的两株牛分枝杆菌(M.bovis C68004和M.bovis N)进行序列比对,对基因突变情况进行统计,分析基因突变对其编码蛋白功能和结构的影响。结果表明:1)临床分离株与本实验室保存的高毒力菌株M.bovis N毒力基因突变位点具有很高的相似性,在检测的29个毒力基因中,50%以上临床分离株与M.bovis N共有25个基因序列一致;2)临床分离株广泛存在的Mb2958、Mb2982C和Mb1553C基因突变PROVEAN评分均低于-2.5,可能改变其编码的酶催化活性,进而影响细胞壁脂类合成;3)Mb0607、Mb0609、Mb0606、Mb2979C和Mb1214基因突变分别造成哺乳动物细胞入侵蛋白、甲基转移酶和酰基转移酶蛋白二级结构改变,或许对细菌的胞内生存产生影响。综上,本研究筛选出8个关键毒力基因,这些基因的突变可能在新疆地区牛分枝杆菌流行菌株的毒力变化中发挥重要作用;同时,本研究揭示了关键毒力基因的突变特征,为进一步研究牛分枝杆菌遗传多样性及进化规律奠定了基础。     

土壤肥力对红壤性水稻土水稻产量和氮肥利用效率的影响

【目的】土壤肥力是红壤性稻田水稻丰产的基础。明确不同肥力对红壤性水稻土作物产量和氮肥利用效率的影响,为红壤性稻田土壤培肥和合理施肥提供科学依据。【方法】选取质地相似的不同肥力水平的红壤性水稻土进行盆栽试验(以土壤有机质的高低代表土壤肥力的高低),利用~(15)N同位素示踪技术研究不同肥力水平(F_L、F_M和F_H分别代表低肥力、中肥力和高肥力,其低、中、高肥力土壤的有机质含量分别为19.9、29.6、38.9 g·kg~(-1))和氮肥水平(N_0、N_(150)和N_(225)分别代表施氮量为0、150和225 kg·hm~(-2),共9个试验处理,分别为F_LN_0、F_LN_(150)、F_LN_(225)、F_MN_0、F_MN_(150)、F_MN_(225)、F_HN_0、F_HN_(150)和F_HN_(225))对水稻产量及其构成、氮肥吸收及其去向的影响。【结果】提升土壤肥力和施氮均能显著提高水稻的有效穗数、产量和总吸氮量。与N_0相比,F_L、F_M和F_H在N_(150)处理下的增产率分别为63%、40%、17%,而在N_(225)处理下的增产率分别为89%、55%和23%。在中、低肥力土壤上,增施氮肥能显著提高水稻产量,而F_HN_(150)和F_HN_(225)处理之间产量无显著差异。~(15)N示踪结果表明,相同施氮量条件下,水稻植株对肥料氮素和土壤氮素的吸收量均随土壤肥力的提高而增加。但是,水稻植株总吸氮量中来自土壤氮素的比例随土壤肥力的提高而增加,而来自肥料氮素的比例则随之降低。增施氮肥会增加水稻吸收肥料氮素的比例,降低其吸收土壤氮素的比例。F_L、F_M和F_H土壤水稻的平均氮肥回收率分别为42%、48%和52%,平均氮肥残留率分别为20%、23%和28%,平均氮肥损失率分别为38%、29%和20%。F_LN_(225)氮肥回收率显著高于F_LN_(150),F_M两个施氮量氮肥回收率无显著差异,而F_HN_(225)的氮肥回收率显著低于F_HN_(150)。提升土壤肥力能显著提高土壤微生物量氮、铵态氮和固定态铵的含量。【结论】提升土壤肥力能显著提高红壤性水稻土的水稻产量以及化肥氮的回收率和残留率,而降低氮肥损失率。在低肥力土壤上适当增加施氮量有利于增加产量和氮肥回收率;而在高肥力土壤上适当降低施氮量在保证较高水稻产量的同时,能够提高氮肥回收率、降低氮肥损失。     

A novel glycoside hydrolase 74 xyloglucanase CvGH74A is a virulence factor in Coniella vitis

Grape white rot is a destructive fungal disease occurring worldwide. Recently, Coniella vitis was identified as the predominant pathogen causing this disease in China. As the periderms of grape shoots are severely degraded by C. vitis, it was speculated that cell wall-degrading enzymes (CWDEs) might play a key role in the pathogenesis of this disease. Therefore, this study aimed to examine the hydrolytic activity of the CWDEs of C. vitis. The results showed that xylanase (Xy) and xyloglucanase (XEG) had high levels of hydrolytic activity both in vitro and in vivo. Furthermore, a high-virulence fungal strain exhibited higher levels of Xy and XEG activities compared with a low-virulence strain. The genome of the fungus was found to harbor two XEG-coding genes CvGH74A and CvGH74B, which belonged to the glycoside hydrolase (GH)74 family. The expression level of CvGH74A was found to be high during pathogen infection. CvGH74A gene deletion mutants were generated using the split-marker method. The deletion of CvGH74A decreased both the hydrolytic activities of XEG and Xy and also the ability of the fungus to infect the grape leaves. No differences in the hyphal growth, morphology of colonies, or conidiation were found between the ΔCvGH74A mutant strains and the wild-type strain. Together, these results suggested that CvGH74A acted as an important virulence factor, and its enzymatic activity might regulate the virulence of the pathogen. This study was novel in reporting that GH74 XEG acted as a virulence factor in C. vitis.     

Enhancement of Arachidonic Acid Through Protoplast Regeneration Production by Mortierella isabellina Mutagenesis

A developed method was used for the enhancement of arachidonic acid production by M. isabellina. An orthogonal, rotatable and central composite design was applied to determine the optimum conditions for protoplast regeneration mutagenesis. The results showed that a commixture enzyme (cellulase and glusulase) at the concentration of 4%, enzymolysis temperature at 30°C and enzymolysis time on 7.5 h were the optimal conditions, in which the lethality of M. isabellina spores was 78.4%. After mutagenesis and re-screenings, M. isabellina mutant Y-69 was obtained. GC analysis showed that the yield of arachidonic acid by Y-69 (2.92 g·L^?1) was 3.56 times higher than that of the wild-type strain (0.82 g · L^?1). Pass generation tests showed that the properties of Y-69 by mutation were readily inherited.     

Study on Mutant Induction of Gladiolus by in vitro Culture of Petals

Petals of Gladiolus 'Rose Supreme' were used to establish regeneration system. Developmental characteristics of petals were observed. A total of 94 plantlets of petal somaclonal line were marked in Mj ...