Productive Traits and Triploid Rate Stability in Triploidy-Induced ‘Haida No. 2’ Strain of the Pacific Oyster, Crassostrea gigas

In order to evaluate the effects of triploidy induction on a selected strain‘Haida No.2’of the Pacific oyster Crassostrea gigas,which is characterized with golden shell color and high growth rate,the growth,survival rate and stability of triploid rate were analyzed at different development stages in the present study.Three different conditions inhibiting the release of polar body Ⅱ or polar body Ⅰ were tested:(A)Cytochalasin-B(CB),0.5mg L^(−1) at 10min post-insemination for 15 min;(B)CB,0.5mg L^(−1)at 15 min postinsemination for 20 min;and(C)CB,0.7mg L^(−1),at 15 min post-insemination for 20 min.The triploidy induction treatments significantly reduced the D-larvae and survival rates at the larvae stage but not at the juvenile and adult stages.Triploid rate dramatically decreased at the larval stage and did not significantly change at the juvenile and adult stages.Regarding the stability of the triploid rate,there was a significant difference between the three treatment groups.Larvae from the treatment A and control groups exhibited higher growth rates in shell height than those from the other two treatment groups at day 27.Triploid juveniles and adults from the treatment A group exhibited a higher wet weight than diploids from the control group and triploids from the other treatment groups.Triploidy induction did not affect the shell color of the progeny.The results obtained in the study demonstrate that triploidy induction has the potential to be used to increase the production of C.gigas variety‘Haida No.2’without modifying its golden shell color.

Selection of Media for Hardwood Cuttings Container Seedling-raising of Triploid Clones of Populus tomentosa

[Objective] The experiment was aimed to select effective and economical media for container seedling of triploid clones of Populus tomentosa that was carried out. [Method] The sandy loam, peat, perlite, vermiculite, riversand, sludge were taken as media of hardwood cutting and survival rate, seedling height were taken as indexes to select media for container seedling of triploid clones of Populus tomentosa. [Result] Different mixedmedia had great influence on survival rates of container seedlings. Taking peat and vermiculite with the proportion of 5∶2 (M10) or peat ,vermiculite with the proportion of 7∶2 (M11) or sandy loam (M1) as media would generate higher cutting survival rate that was higher than 90.0%. There were significant differece in height increments of container seedlings. Taking sandy loam, peat and vermiculite with the proportion of 6∶2∶2(M5)or sandy loam (M1), seedling height of 60-days the seedling was over 37.0 cm. [Conclusion] According to cost analysis of nursery medium, the optimum medium for hardwood cuttings container seedling-raising of triploid clones of Populus tomentosa was sandy loam.

Establishment of in Vitro Regeneration System of Triploid Chinese White Poplar

初步建立了三倍体毛白杨 (PopulustomentosaCarr.)组织培养再生系统。组织培养时 ,外植体最好选用在弱光下从休眠芽上萌发的幼芽或嫩茎 ,经HgCl2 溶液表面消毒后 ,接种到大量元素减半、含 0 .2 5mg/L吲哚乙酸和 1.0 0mg/L玉米素的MS培养基上。试管苗的生根可采用含 0 .2 5mg/L吲哚丁酸和 10mg/L维生素B1的MS培养基。试管苗茎段的分化依无性系和外源激素条件的不同而异。无性系B19的茎段较易分化 ,而无性系B30 4和B331分化较难。最适的分化培养基为大量元素减半、含 0 .0 5～ 0 .10mg/L吲哚乙酸和 1.0

Establishment of High Frequency Regeneration System of Populus tomentosa

The establishment of high frequency regeneration system is a foundation for Agrobacterium mediated genetic transformation. In this work, several important factors influencing the efficiency of regeneration of plants, such as concentration of plant growth regulators, leaf explant orientation, leaf growth sequence and leaf segment, were studied. The results indicated that the differentiation rate of adventitious shoots was 90% on basal MS medium only supplemented with 1 5?mg·L -1 BA (6 benzyladenine) and reached the highest level(95%) when 1 0?mg·L -1 BA and 0 3?mg·L -1 NAA (naphthaleneacetic acid) were added to MS medium. 90% of differentiation rate of adventitious roots were obtained when 0 3?mg·L -1 NAA was only added to MS medium. It was also found that more adventitious shoots were regenerated from the lower segment of leaf (with petiole) than the other segments, the number of adventitious shoots decreased from top to base of leaf growth sequence and the percentage of adventitious shoot induction with adaxial side downward was higher than that with adaxial side upward.

恩诺沙星对湘云鲫(Triploid crucian carp)组织转氨酶(GPT、GOT)活性的影响

采用胸腔注射法,研究了恩诺沙星对湘云鲫(Triploid crucian carp)组织谷草转氨酶(GOT)和谷丙转氨酶(GPT)活性的影响。结果显示,给药后5 d,各试验组GPT、GOT酶活性均升高,与对照组之间呈显著差异水平(P<0.05);给药后10 d,各浓度组GPT酶活性有所降低,而GOT酶活性则继续升高,鳃的GOT酶活性达到了最大值;给药后15、20 d,各浓度组GPT酶活性继续下降,而肝脏、肾脏GOT酶活性在给药后15 d时达到最大值,之后开始下降;给药后25 d,鳃的GPT酶活性除200 mg/kg组外都恢复至正常水平,而肝脏、肾脏各浓度组的GPT酶活性显著低于对照组(P<0.05),肝脏的GOT酶活性恢复到了正常水平,而鳃和肾脏的GOT酶活性略高于对照组水平。

细胞松弛素B和低温诱导华贵栉孔扇贝三倍体的产生(英文)

分别用细胞松驰素B（C．B）（浓度为0．5mg／L）和低温（10℃）处理诱导华贵栉孔扇贝Chlamysnobilis产生三倍体．抑制第一极体的三倍体诱导率达到44．1％和31．8％；而抑制第二极体的三倍体诱导率分别为90．2％和72．7％（胚胎初期检查）．随着C．B浓度的增加，孵化率下降；而随着温度的降低和休克时间的延长，非整倍体增多．

Genetic study on seed coat characteristics in multiploid watermelon

[ Objective] This study is to provide basis for the genetic improvement of triploid watermelon cultivars from seed coat characteristiscs by analyzing its genetic properties. [ Method ] Genetic effects of seed coat characteristics and their related characteristics in tdploid watermelon were analyzed by additive dominant genetic model based on 36 F1 hybrids which were crossed by six female parents of four ecotypes and six male parents of same ecotype according to North Carolina II crossing design. [ Result] The seed coat number characteristics was controlled by additive effects, dominant effects and error variance of genetic together. The additive effects of the length and width of abortive seed coat were significant, and the error variance was significant or highly significant. It was observed that there was a significant negative correlation of additive effects between the seed coat number and fringe soluble sugar. Also there was a significant negative correlation of dominant effects between lycopenes, dry matter and the seed coat number. However, there was a highly significant positive correlation in additive effects between the length and width of abortive seed coat. [ Conclusion ] The results provide guidance for genetic breeding of triploid watermelon cultivars.

Discovery of Triploid Allium macrostemon Bunge in Laizhou City of Shandong Province

[Objective]The study was to analyze the karyotype of Allium macrostemon Bunge in Laizhou City of Shandong Province.[ Method] The root tip of A. macrostemon was pretreated with 8-hydroxyquinoline solution, fixed, dissociated and stained for preparing the glass slide to reveal the chromosome number via the microscopic examination; the sparse cells with good chromosome morphology were photographed under microscope. [ Result ]Allium macrostemon Bunge in Laizhou City introduced in this study was tdploid; its somatic chromosome number was 24 and karyotype formula was K（2n） =3x=24m（2SAT） ＋ 1Bs, thus the karyotype belongs to 1A type. One of the chromosome No. 3 contained satellite, and chromosome deletion may be existed in one of the chromosome No. 5. In addition, B chromosome was observed in some cells.[ Conclusion] This introduction of triploid A. macrostemon found in China was the first time.

Genetic Control of Air-Dried Wood Density, Mechanical Properties and Its Implication for Veneer Timber Breeding of New Triploid Clones in Populus tomentosa Carr

The wood samples of 9 triploid clones of Populus tomentosa Carr. taken from a 9 year old clonal test site were analyzed in order to investigate the genetic variation of wood properties, including air dried wood density and some mechanical properties. The results showed that significant or extremely significant difference in air dried wood density and the mechanical properties existed among the clones, this means these wood properties were under moderate or strong genetic controls and could be improved by genetic manipulations. The radial and vertical variation patterns of air dried wood density were also studied and the results were found to coordinate with other previous research results. The vertical variation patterns of most mechanical properties within the individual tree also conformed to the general wood theories except the modulus of elasticity and cross section hardness. Among the mechanical properties, modulus of elasticity (MOE) and tangent section hardness were under strong genetic control, with the clonal repeatabilities being 0 90 and 0 80, respectively. However, the clonal repeatabilities of other mechanical properties under study were a little lower than above two indexes. Genetic correlation analysis indicated that super clonal selection and breeding for veneer timber could be realized through indirect selection of wood density and form indexes.

In Vitro Development and Germination of Immature Embryos of Sweet Oranges Parson Brown and Pineapple(Citrus sinensis)

Oranges are classified as the most traded fruit variety worldwide.The diversification of the varietal profile of this group is among the objectives of the Citrus breeding program.So,the creation of new triploid hybrids via immature embryos rescue,can lead to an increase in gene pool and produce seedless varieties.In addition,flow cytometry was successfully used for analyzing ploidy levels and genetic origin of triploid plants.The present study was aimed at optimizing immature embryos in vitro germination and plantlet development based on medium chemical composition of the two orange varieties,Pineapple and Parson Brown.When the fruit gets matured,they are harvested and the seeds extracted are classified according to their size.Only those which are small are cultured in a sterile Murashige and Tucker(MT)basal medium.The different concentrations of growth regulators are put to the test in order to gain the best medium for seeds development.M1(MT+1 mg/L gibberellic acid(GA3)),M2(MT+1 mg/L kenitin+0.5 mg/L 6-benzylaminopurine(BAP)+0.1 naphthalene acetic acid(NAA)),M3(MT+25 mg/L adenine sulfate(ADS)),M4(MT+0.5 mg/L kenitin+0.5 mg/L BAP+1 mg/L GA3).Percentage of germinated embryos(PGE)is the highest in M3 with 100%and 90%for both Pineapple and Parson Brown varieties,respectively.In addition,the germination interval average(GIA)is shorter in M1 for both varieties,which takes 6 d and 7 d,respectively.As regard to speed seedling growth(mm/week),both varieties undergo significant change in all four mediums.Similarly,the highest rate of acclimatization for the Pineapple and Parson Brown varieties is 80%and 90%in M1.In general,the smaller the embryos are,the more sensitive they are to culture medium composition.It is therefore essential to optimize the medium components in order to stimulate their in vitro development.Therefore,M1(MT+1 mg/L GA3)remains the best to promote short term good germination and produce a better rate of acclimatization.

Cytological Basis of Polyspermic Fertilizationin An Artificial Multiple Triploid Carp1

The fine structure of the micropyle and the initial stages of sperm entry into the egg of the artificial multiple triploid carp were observed by means of scanning electron microscopy. Only one micropyle at the animal pole of the mature egg could be found. The aperture of the outer opening of the micropylar canal was only wide enough to admit one spermatozoon to pass through. A series of ordered sequence of motile events after fertilization were observed, which including that the sperm attached to the surface of the mature eggs and passed through micropylar canal, cottonlike substances closed up the micropylar canal, cortical vesicles underwent exocytosis and digesting, the vitelline envelope was transformed into fertilization envelope and so on. In addition, a preliminary discussion on the mechanism of polyspermic fertilization in the artificial multiple triploid carp was described in this paper.

Polyploidy induction by hydrostatic pressure shock and embryo development of sea cucumber Apostichopus japonicus

The manipulation of the chromosome set for commercially valuable marine animals is important for enhancing aquacultural production. In this study, triploid and tetraploid sea cucumber Apostichopus japonicus were induced by hydrostatic pressure shock, and the conditions of appropriate induction were tested with different starting times, and hydrostatic pressure intensities and durations. The highest rate of triploid induction reached 20% and that of tetraploid was 60%. In consideration of the survival rate and hatch rate, the appropriate treatment for triploid was 55 Mpa of hydrostatic pressure for 5 rain at 55 min after fertilization （a.f.）, while for tetraploid it was 60 Mpa for 5 rain at 61 min a.f. The triploid of the sea cucumber could survive through the pelagic larval stage and attachment stage, and develop like the control group of the experiment. The tetraploid, however, could not survive the attachment stage.

Comparison of Taste Components between Triploid and Diploid Oyster

The main taste components of triploid and diploid oyster (Crassostrea gigas) were compared. Free amino acids, inosine monophosphate, succinate, trimethylamine oxide and betaine in fresh and boiled extractives were analyzed. The protein, lipids, glycogen, moisture and ash, which may affect the flavour, were evaluated. In boiled extractives, the amino acids were 394.1 mg(100 g) -1 in diploid and 183.5 mg(100 g) -1 in triploid. However, in fresh oyster extractives, they were 320.0 mg(100 g) -1 and 147.3 mg(100 g) -1 respectively. The inosine monophosphate in triploid was 44% more than that in diploid, and a little difference existed in the content of trimethylamine oxide between them. The contents of these taste components were the basis for taste flavour pattern determination.

Natural Homologous Triploidization and DNA Methylation in SARII-628,a Twin-seedling Line of Rice (Oryza sativa)

A total of five pairs of diploidotriploid twin-seedlings （a diploid seedling and a triploid seedling emerged from a grain） were selected out from 4500 pairs of seedlings from SARII-628, a twin-seedling rice line. SSR analysis indicated that no difference between the diploid seedling and corresponding triploid seedling in a twin-seedling was found at the 310 loci, indicating that there was no obvious change in DNA primary structure. A modified AFLP technique ＇MSAP （methylation-sensitive AFLP）＇ was used to analyze methylation mutation. Although no methylation mutation was noted among the five diploids, 29 methylation mutation loci were found from the corresponding triploids. This suggested that methylation mutation happened rapidly on Mogeneration after natural homologous triploidization. The mutations were classified into 10 types, including 3 increased types, 3 decreased types and 4 undecided types of methylation-degrees. The bands of 22 loci were sequenced and then those sequences were searched through website. The result showed that the methylation mutation involved into the whole rice genome and the 12 pairs of chromosomes. The mutation trend was site-related and there were different mutation loci for different triploids, which foretold that SARII-628 would have different evolution fates after natural homologous triploidization.

PRELIMINARY STUDY ON CHROMOSOMES OF INDUCED TRIPLOID IN THE RED SEA BREAM, PAGROSOMUS MAJOR

Triploidy red sea bream were induced by cold shock techniques (0 3℃) in Qingdao in April 1994, May 1995 and May 1996. Normal diploidy and triploidy chromosome metaphases were produced by chromosome spreads from the gastrula. Counts of 104 chromosome metaphases of normal diploid showed each of them consisted of 2 acrocentric (st) and 46 telocentric (t) chromosomes. Based on the relative lengths and arm ratios, the 48 chromosomes were matched into 24 pairs. Counts of 107 chromosome metaphases of induced triploid showed that each metaphase consisted of 3 acrocentric (st) and 69 telocentric (t) chromosomes. The 72 chromosomes were easily matched into three sets of chromosomes, based on the relative lengths and arm ratios.

Seasonal Variation of the Glycogen Enzyme Activity in Diploid and Triploid Pacific Oyster Gonad During Sexual Maturation

The glycogen content and the activities of two key enzymes in glycogen metabolism, glycogen phosphorylase and gly- cogen synthetase, in the gonad of diploid and triploid Pacific oysters (Crassostrea gigas) were compared during maturation. The glycogen content in the gonad of diploids decreased with gametogenesis (by 85.7%), but the glycogen content in the gonad of trip- loids did not vary significantly. Activity of glycogen phosphorylase (GP) in the gonad of diploids decreased with gametogenesis (by 55.5%), while GP activity of triploids did not vary significantly during maturation. Activity of glycogen synthetase (GS) in the gonad of diploids increased slightly with gametogenesis, reaching a peak in June. Activity of GS declined sharply from June to July, which might be due to gonad spawning. GS activity of triploid oysters in spawning time (July and August) was significantly higher than that in other months, which might be explained with a ‘compensating’ mechanism for the higher glycogen content in triploids.

Transcriptomic Analysis of Pacific Oyster(Crassostrea gigas) Zygotes Under Hypotonic Triploid Induction

Polyploid breeding is widely used in various marine species. Low salinity treatment is an effective method of inducing triploid of bivalve mollusks. In this study, RNA-seq was performed to determine genes and pathways involved in hyposaline adaption and cell division of Pacific oyster(Crassostrea gigas) zygotes, trying to better understand the possible molecular mechanism of hypo-osmotic induction. A total of 26965 unigenes were generated in the de novo assembly of clean Illumina reads with an average length of 934 bp and N50 of 1721 bp. Of 3024 differentially expressed genes(DEGs), 2501 were up-regulated and 523 were downregulated. GO(Gene Ontology) annotation and KEGG(Kyoto Encyclopedia of Genes and Genomes) pathway analysis of these DEGs revealed that these DEGs participate a variety of biological processes including osmoregulation, cytoskeleton organization, cell survival and death, and substantially modulate cell proliferation and embryonic development. In summery, RNA-seq methodology was applied for the first time to demonstrate hypotonic-induced transcriptomic alteration in oyster zygotes. Our findings not only interpreted the relatively high mortality of induced larvae, but also provided a valuable reference for further investigations on the mechanism of hyposaline induction, thus should aid to the application of low salinity in triploid induction in large scale aquaculture in future.

Chromosome Behavior of Heat Shock Induced Triploid in Fenneropenaeus Chinensis

Triploidy was induced in Chinese shrimp Fenneropenaeus chinensis by 30±0.5℃heat shock treatment (initiated at 20 min after fertilization) for 10 min to inhibit the release of PB2 at 18.0℃. The highest triploid rate obtained was 84.5% in nauplius stage. The effect of heat shock treatment on meiosis and cleavage of eggs was investigated in this work aimed to establish efficient procedures for triploid induction and to gain understanding of the mechanism of triploid production. Three pronuclei that could be observed in the treated eggs under fluorescence microscope developed into triploid embryos. Some abnormal chromosome behavior was observed in heat shocked eggs.

Integration of Ru/C and base for reductive catalytic fractionation of triploid poplar

Lignin,which is the most recalcitrant component of lignocellulosic biomass,is also the most abundant renewable aromatic resource.Herein,reductive treatment of triploid poplar sawdust by the integration of catalytic Ru/C and a base,which afforded high yields of phenolic monomers from the lignin component and a solid carbohydrate pulp,is reported.The introduction of Cs_(2)CO_(3) led to the generation of C2 side‐chained phenols through the cleavage of C_(β)–O and C_(β)–C_(γ) bonds inβ–O–4 units in addition to C3 side‐chained phenols;the relationship between C2 and C3 was dependent on the base dosage.The reaction conditions,including base species,temperature,time,and H_(2) pressure,were optimized in terms of phenolic product distribution,delignification degree,and carbohydrate retention.The carbohydrate pulps generated from reductive catalytic fractionation in the presence of Cs_(2)CO_(3) were more amenable to enzymatic hydrolysis,indicating that this treatment of biomass constituted the fractionation of biomass components together with the breakdown of biomass recalcitrance.

Microsatellite Analysis of Genetic Diversity Between Loach with Different Levels of Ploidy

We used microsatellite markers to investigate the genetic parameters of three different polyploidy populations of Misgurnus anguillicaudatus from Honghu City, Hubei Province. Polyacrylamide gel electrophoresis banding patterns of diploid（2n）, triploid（3n） and tetraploid loaches（4n） were analyzed with Pop Gen software. A total of 68 alleles were obtained from seven microsatellite loci and the polymorphism information content（PIC） indices were all above 0.5. The average expected mean heterozygosity values（He） were 0.8420, 0.7186 and 0.8521; the average observed mean heterozygosity values（Ho） were 0.9674, 0.9785 and 0.8928; and the HardyWeinberg P values were 0.3078, 0.3151 and 0.3762, for diploid, triploid and tetraploid individuals, respectively. The results indicated that the three populations were highly polymorphic, with no deviations from Hardy-Weinberg equilibrium observed at all the seven microsatellite loci. This indicated a high level of genetic diversity within the populations. A cluster analysis diagram showed that the shortest genetic distance was between diploid and tetraploid loaches and they shared a close phylogenetic relationship. The triploid and tetraploid individuals had the most distant phylogenetic relationship.