Early Embryo and Larval Development of Inviable Intergeneric Hybrids Derived from Crassostrea angulata and Saccostrea cucullata

To detect the intergeneric hybridization between the oyster Crassostrea angulata and Saccostrea cucullata coexisting along the southern coast of China, reciprocal crosses were conducted between the two species. Barriers for sperm recognizing, binding, penetrating the egg, and forming the pronucleus were detected by fluorescence staining. From the results, although fertilization success was observed in hybrid crosses, the overall fertilization rate was lower than that of intraspecific crosses. A large number of hybrid larvae died at 6–8 d after hatching, and those survived could not complete metamorphosis. C. angulata ♀× S. cucullata ♂ larvae had a growth rate similar to that of the maternal species, whereas S. cucullata ♀× C. angulata ♂ larvae grew the slowest among all crosses. Molecular genetics analysis revealed that hybrid progeny were amphimixis hybrids. This study demonstrated that hybrid embryos generated by crossing C. angulata and S. cucullata could develop normally to the larval state, but could not complete metamorphosis and then develop to the spat stage. Thus, there is a post-reproductive isolation between C. angulata and S. cucullata.

Phytogenetic Studies on Intergeneric Hybridization Between Brassica napus and Matthiola incana

The intergeneric hybridization between Brassica napus L. cv. oro and Matthiola incana (L.) R. Br. was carried out to study the phytogenetic peculiarities of the hybrid plants. In order to improve the oil quality of rape seed, ovaries of B. napus pollinated with pollen of M. incana, were cut off and inoculated onto MS media supplemented with various plant hormones at the 7th day after pollination. Two mature embryos were obtained from 750 pollinated ovaries cultured in vitro. The percentage of seed set was 0.26%. The mature embryos were transferred onto the MS media supplemented with 2.0 mg/L 6-BA + 0.1 mg/L NAA, and a compound bud was produced later. The compound bud was then cut into a number of single buds, which were transferred onto fresh media. Twenty-two plantlets in vitro were developed from the above single buds. The hybrid plants (F-1) were basically intermediate between the two parents in many,characters with a few showing hybrid vigor. The fertility of hybrid plants (F-1) was poor. Cytological studies revealed that the hybrid plants (F-1) were mixoploid. in nature. The chromosome number of many somatic cells was 2n = 26, which was the sum of the chromosome number of the two parents. The chromosome number of other somatic cells was 2n = 38, similar to that of B. napus. The hybrid offspring (F-2) from the selfed hybrid plants (F-1) showed polymorphism. Among the hybrid offspring (F-2) some were nearly matroclinous and fertile, similar to B. napus. some others were intermediate between the two parents and less fertile, and a few were poorly developed and nearly infertile. From the hybrid offspring a few plants with improved seed-oil quality were obtained.

Preliminary Studies on the Intergeneric Hybridization Between Brassica napus with OguCMS and Orychophragmus violaceus

In the cross between B. napus with OguCMSOrychophragmus violaceus, pre-fertilizatiocompatibility was observed. An intergeneric hybrid was successfully obtained by usinovary culture. The hybrid status was confirmed by cytology, morphology, fertilityisozyme and miosis analysis. The seed setting rate was 0.0072%. An amphidiploid of thhybrid was also produced by the use of chromosome-doubling method with 0.1% colchicinetreatment. Obtaining this hybrid and its amphidiploid has important theoretical anutilization value.

Varietal differences in photosynthetic characters and chlorophyll fluorescence induction kinetics parameters among intergeneric progeny derived from Oryza×Sorghum, its parents, and hybrid rice

A comparative study on the photosynthetic parameters among intergeneric progenies derived from \%Oryza sativa\% L.×\%Sorghum vulgare\% L., its maternal parent Gui 630 and commercial 3\|line hybrid rice Shanyou 63 in pot experiment in greenhouse was conducted. The morphological and photosynthetic characters of canopy leaves and chlorophyll fluorescence kinetic parameters including \%F\%\-v/\%F\%\-m, \%F\%\-v/\%F\%\-0, photochemical quenching coefficient and non\|photochemical coefficient of canopy leaves of 3 varieties were measured. The results showed the progeny, Yuanyou 1, derived from an intergeneric cross of rice and sorghum possesses better canopy spatial architecture with thicker, heavier and bigger canopy leaf than its maternal parent Gui 630. Higher photosynthetic rate due to higher chlorophyll content, higher primary energy transformation efficiency, potential of PSII and non\|photochemical quenching coefficient (\%q\%\-E) were also measured in Yuanyou 1. These explain partly why the intergeneric progeny has higher biomass production, and better tolerance to adverse conditions and higher field yields even under stress conditions.

Dominance of Brassica and No Effects of Raphanus in Mature Seed Production in Intergeneric Hybrid between Brassica rapa ssp. Pekinensis and Raphanus

We succeeded in producing mature seed from a line of Brassica rapa ssp. pekinensis that had been hybridized with Raphanus sativus var. major. Our focus was on dominance of B. rapa ssp. pekinensis;radish (R. sativus var. major) had no influence. Marker tests for similarity showed that the original CR291M-64 x HwiM-2 hybrid was an inbred CR291M-64, rather than a genuine cross;this appears to have resulted from weak self-incompatibility in this strain. The plants from the mature seed bloomed with reddish flowers differently shown up to present. The intergeneric hybrid between Brassica inbred and Raphanus hybrid was very weak in strength compared to the Brassica inbred which was self-pollinated even though the cause of the weak was not identified. The hybrids between Brassica hybrid, dominant and elite recessive, and Raphanus can be developed in large quantities using mature hybrid seed without resorting to ovule culture techniques.

Analysis of Disequilibrium Hybridization in Hybrid and Backcross Progenies of Saccharum officinarum × Erianthus arundinaceus

Erianthus arundinaceus is an important, closely related genus of Saccharum officinarum L. It is therefore important to understand how the chromosomes are transmitted when it hybridizes with sugarcane. The hybrids and backcross progenies of S. officinarum and E. arundinaceus and their parents were used for Karyotype analysis and to study the law of chromosome transmission. The results showed that the somatic chromosome number of both of the E. arundinaceus Hainan92-105 and Hainan92-77 were 2n = 60 = 60sm, belonging to type 1 A, and the BC1 YC01-21 was 2n = 104 = 100m ＋ 4sm, belonging to type 2C. The other six tested clones belonged to type 2B. The both F1s YC96-66 and YC96-40 that originated from Badila （2n = 80 = 70m ＋ 10sm） with E. Arundinaceus were 2n = 70 = 68m ＋ 2sm, which suggests an n ＋ n transmission. The cross between YC96-66 （female parent） and CP84-1198 （male parent, 2n = 120 = 114m ＋ 6sm） also followed the same genetic law and the somatic chromosome number of their progeny, YC01-3 （2n = 105 = 95m ＋ 10sm）. The cross derived from YC96- 40 （female） and CP84-1198 （male）, YC01-21 had 2n = 104 = 100m ＋ 4sm chromosomes, following the same genetic law of n ＋ n. However, YC01-36 had 2n = 132 = 130m ＋ 2sm chromosomes, which suggests a 2n ＋ n chromosome transmission. It can be inferred that the inheritance of chromosomes was very complex in the BC1. The difference in chromosome number between clones was as high as 28. This could be explained by the 2n ＋ n transmission of chromosomes. In addition, as there was not be a regular number of haploids, this phenomenon is termed as disequilibrium hybridization.

MiRNA identification,characterization and integrated network analysis for flavonoid biosynthesis in Brassicacoraphanus

Brassica rapa(AA,2n=20)and Raphanus sativus(RR,2n=18)are popular vegetables containing multiple secondary metabolites,such as flavonoids and anthocyanins.Nevertheless,fewstudies on miRNA expression profiles and potential regulatory network of flavonoid metabolism have been reported in the intergeneric hybrid of B.rapa and R.sativus(Brassicacoraphanus,AARR,2n=38).Here,themiRNAs of Brassicacoraphanus and its parents were first identified by small RNA sequencing.It was revealed that 315 conserved miRNAs and 193 novel miRNAs were found in Brassicacoraphanus and its parents.Moreover,the miRNA expression profiles were characterized and 204 miRNAs with differential expression were obtained.Further analysis showed that 184 miRNAs were non-additively expressed in Brassicacoraphanus.At last,the potential regulatory network of flavonoid metabolism in Brassicacoraphanus was analyzed by multi-omics data,suggesting possible roles of miR858a and miR157a in regulating flavonoid synthesis pathway in Brassicacoraphanus.Taken together,this study provides useful resources for probing globalmiRNA expression patterns in Brassicacoraphanus,which facilitates regulatory mechanism research on the biosynthesis pathway of secondary metabolites in Brassicacoraphanus.

Intergeneric Somatic Hybridization Between Brassica napus L. and Sinapis alba L.

: Electrically induced protoplast fusion was used to produce somatic hybrids between Brassica napus L. and Sinapis alba L. Seven hybrids were obtained and verified by the simple sequence repeat and cleaved amplified polymorphic sequence analysis of the gene fael, indicating that the characteristic bands from S. alba were present in the hybrids. The hybridity was also confirmed by chromosome number counting because the hybrids possessed 62 chromosomes, corresponding to the sum of fusion-parent chromosomes. Chromosome pairing at meiosis was predominantly normal, which led to high pollen fertility, ranging from 66% to 77%. All hybrids were grown to full maturity and could be fertilized and set seed after self-pollination or back-crosses with B. napus. The morphology of the hybrids resembled characteristics from both parental species. An analysis of the fatty acid composition in the seeds of F1 plants was conducted and the seeds were found to contain different amounts of erucic acid, ranging from 11.0% to 52.1%.

PCR-RFLP and AP-PCR of rbcL and ITS of rDNA Show That × Taxodiomeria peizhongii （ Taxodium × Cryptomeria） Is not an Intergeneric Hybrid

Taxodiorneria peizhongii Z. J. Ye, J. J. Zhang et S. H. Pan was regarded as a new Intergenerlc hybrid between Taxodlum mucronatum Tenore （as the female donor） and Cryptomeria fortunei Hoolbrenk ex Otto et Dletr （as the male donor）. To confirm the authenticity of the intergeneric hybrid, we analyzed the rbcL gene and the Internal transcribed spacer （ITS） of 26S-18S ribosomal RNA gene of the three species using polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP） and arbitrarily primed PCR （AP-PCR）, and ob- tained the following results：i） Taxodiomeria peizhongii had the same RFLP maps of the rbcL gene and the ITS as Taxodlum mucronatum, but was different from C. fortunei; II） a 311-bp PCR amplification product was obtained In C. fortunei by AP-PCR of ITS, but was not found in Taxodiomeria peizhongii. Our results have demonstrated that C. fortunei did not provide any genome for Taxodiomeria peizhongii, Implying that T. peizhongii Is not an Intergenerlc hybrid between the two species.