Confirmation of Pearl Millet-Napiergrass Hybrids Using EST-Derived Simple Sequence Repeat (SSR) Markers

Prospects for deploying perennial grasses that are currently considered leading candidates for dedicated energy crops over large acreages are debatable because of several limitations, including vegetative propagation or small seed size, low biomass production during the first growing season, and incomplete assessments of crop invasiveness risk. Pearl Millet-Napiergrass hybrids (“PMN”;Pennisetum glaucum [L.] R. Br. × P. purpureum Schumach.), in contrast, are large-seeded, sterile feedstocks capable of high biomass production during establishment year. Novel methods are warranted for confirmation of PMN hybrids, as traditional morphological observations can be inconclusive and chromosome number determination using cytological methods is laborious and time consuming. Six putative PMN lines were produced in this study, and 10 progeny from each line were evaluated using morphological traits, seed fertility, flow cytometry, and expressed sequence tag-simple sequence repeat (EST-SSR) markers. All putative hybrid lines were sterile and failed to produce seed. The PMN hybrids could not be distinguished from either parent using flow cytometry due to highly similar nuclear genome DNA contents. A number of paternal napiergrass-specific EST-SSRs were identified for each PMN line, and four paternal-specific EST-SSRs conserved across all napiergrass accessions were selected to screen the putative PMN hybrids. These EST-SSRs confirmed that all F1 individuals analyzed were PMN hybrids. The use of paternal-specific markers therefore provides a valuable tool in the development of both “Seeded-yet-Sterile” biofuel PMN feedstocks and additional PMN cultivar-and parental species-specific markers.

Cytological Analyses on Development of Male and Female Gametophytes in an Interspecific Hybrid F_1 from Cucumis hystrix Chakr. × Cucumis sativus L.

An interspecific hybrid F1 of Cucumis hystrix Chakr. x Cucumis sativus L. （NC4406） was used to establish the developmental sequence and to characterize the male and female gametophytes at cytological level for further understanding of the phylogenic relationship and the mechanism of fertility or sterility in the interspecific hybrid F1 The development of male and female gametophytes was studied through meiotic analysis and paraffin section observation technique, respectively. Meanwhile, the fertility level was assessed through hybrid F1 backcrossing to cultivated cucumber 4406. Variable chromosome configurations were observed in the pollen mother cells （PMCs） of hybrid F1 at metaphase Ⅰ , e.g., univalents, bivalents, trivalents, quadravalents, etc. At anaphase Ⅰ and Ⅱ, chromosome lagging and bridges were frequently observed as well, which led to the formation of polyads and only a partial number of microspores could develop into fertile pollen grains （about 23.3%）. Observations of the paraffin sections showed numerous degenerated and abnormal embryo sacs during the development of female gametophytes, and only 40% of the female gametophytes could develop into normal eight-nuclear megaspore. On an average, 22.8 and 6.3 seeds per fruit could be obtained from the reciprocal backcross. The interspecific hybrid F1 of C. hystrix x NC4406 was partially fertile; however, the meiotic behaviors of hybrid F1 showed a high level of intergenomic recombination between C. hystrix and C. sativus chromosomes, which indicated that it plays an important role for introgression of useful traits from C. hystrix into C. sativus.

Interspecific Hybridization between <i>Arisaema sikokianum</i>and <i>A. serratum</i>(Araceae) Confirmed through Nuclear and Chloroplast DNA Comparisons

A morphologically intermediate plant between Arisaema sikokianum Franch. et Sav. and A. serratum (Thunb.) Schott has been newly found in Kochi Prefecture, Shikoku, Japan. The putative hybrid has the intermediate morphological characteristics of the parental species. Molecular analysis using PCR-RFLP of internal transcribed spacer (ITS) in nuclear DNA (nrDNA) indicates that the putative hybrid has a combined pattern of the two putative parent species. Moreover, the sequence result of chloroplast DNA (cpDNA) of the putative hybrid was identical to that of A. sikokianum. These results suggest that the putative hybrid is a hybrid between A. sikokianum and A. serratum and that it was formed by interactive gene exchanging via pollens from A. serratum to A. sikokianum. It is the first record of a hybrid between A. sikokianum and A. serratum.

Ways of Developing Plants in Interspecific Hybridization of Cotton

It is known,that there are various barriers to fertilization,development of embryos,and endosperm because of different number of chromosomes in parents in the interspecific hybridization of cotton.Thus the factors providing normal cell fission of a germ and endosperm are necessary.It is necessary to culture embryos in vitro on the artificial environments containing various phytohormones,or

Fiber Quality Improvement by Interspecific Hybridization in Upland Cotton

In order to transfer genes for extra fiberstrength to existing cotton cultivars,weintroduced 207 interspecific materials obtainedby outcrossing where G. hirsutum,G.barbadense,G.sturtianum,G.thurberi,G.harknessii and G.somalense etc.were involved asparents respectively.Under stringent

Development of a set of SSR markers for genetic polymorphism detection and interspecific hybrid jute breeding

Corchorus capsularis(white jute) and C. olitorius(dark jute) are the two principal cultivated spedes of jute that produce natural bast fiber of commercial importance. We have identified 4509 simple sequence repeat(SSR) loci from 34,163 unigene sequences of C. capsularis to develop a non-redundant set of 2079 flanking primer pairs. Among the SSRs, trinudeotide repeats were most frequent(60%) followed by dinucleotide repeats(37.6%). Annotation of the SSR-containing unigenes revealed their putative functions in various biological and molecular processes, including responses to biotic and abiotic signals. Eighteen expressed gene-derived SSR(eSSR) markers were successfully mapped to the existing single-nucleotide polymorphism(SNP) linkage map of jute,providing additional anchor points. Amplification of 72% of the 74 randomly selected primer pairs was successful in a panel of 24 jute accessions, comprising five and twelve accessions of C.capsularis and C. olitorius, respectively, and seven wild jute spedes. Forty-three primer pairs produced an average of 2.7 alleles and 58.1% polymorphism in a panel of 24 jute accessions. The mean PIC value was 0.34 but some markers showed PIC values higher than 0.5, suggesting that these markers can efficiently measure genetic diversity and serve for mapping of quantitative trait loci(QTLs) in jute. A primer polymorphism survey with parents of a wide-hybridized population between a cultivated jute and its wild relative revealed their efficacy for interspecific hybrid identificatioii. For ready accessibility of jute eSSR primers, we compiled all information in a user-friendly web database, JuteMarkerdb(http://jutemarkerdb.icar.gov.in/) for the first time in jute.This eSSR resource in jute is expected to be of use in characterization of germplasm, interspecific hybrid and variety identification, and marker-assisted breeding of superior-quality jute.

Comparative Embryological Studies on Infertility of Interspecific Hybridizations Between Oryza sativa with Different Ploidy Levels and O. officinalis

As maternal parents, diploid （L202-2x） and autotetraploid （L202-4x） of Oryza sativa cv. L2O2 were crossed with O. officinalis. Embryo development and fertilization in these two crosses were comparatively studied. There were no mature hybrid seeds obtained because all the hybridized spikelets died 30 days after pollination. The main reasons for no seed set were abnormal fertilization and development of the embryos and endosperms in the interspecific hybrids. There were doublefertilization, egg cell single-fertilization and non-fertilization in these crosses. Although 59.45% and 54.87% of hybrid embryos produced in the crosses of L202-2x/O. officinalis and L202-4x/O. officinalis, respectively, hybrid embryos ceased to develop or degenerated and plenty of free endosperm nuclei were in disaggregating state without developing cellular endosperms three days after pollination. Besides, some embryological differences in these two crosses were found, that is, the rate of double-fertilization and total rate of doubleand single-fertilization in L202-2x/O. officinalis were higher than those in L202-4x/O. officinalis. The embryo and endosperm of hybrids developed more slowly, and embryos and free endosperm nuclei were more severely degenerated in L202-4x/O. officinalis than in L202-2x/O. officinalis. Five days after pollination, a few of embryos in L202-2x/O. officinalis developed into pear-shaped ones, however, embryos in L202-4x/O. officinalis were all degenerated. Therefore, it is more difficult to obtain interspecific hybrids by wide crosses between autotetraploid of O. sativa and O. officinalis.

Molecular Cytogenetic Analysis of Spontaneous Interspecific Hybrid Between Oryza sativa and Oryza minuta

Genomic in situ hybridization (GISH) is a powerful tool to characterize parental chromosomes in interspecific hybrids, including the behaviour of autosynapsis and chromosome pairing. It was used to distinguish the chromosomes of Oryza sativa from wild species in a spontaneous interspecific hybrid and to investigate the chromosome pairing at metaphase I in meiosis of the hybrid in this study. The hybrid was a triploid with 36 chromosomes according to the chromosome number investigated in mitosis of root tips. During metaphase I of meiosis in the hybrid, less chromosome pairing was observed and most of the chromosomes existed as univalent. Based on GISH and FISH (Fluorescent in situ hybridization) analyses, the chromosomes of the hybrid were composed of genomes A, B and C. Thus, it was believed that the hybrid was the result of natural hybridization between cultivated rice and wild species O. minuta which was planted in experimental fields.

Progress on Transferring Elite Genes from Non-AA Genome Wild Rice into Oryza sativa through Interspecific Hybridization

The progress of research on transferring elite genes from non-AA genome wild rice into Oryza sativa through interspecific hybridization are in three respects, that is, breeding monosomic alien addition lines （MAALs）, constructing introgression lines （ILs） and analyzing the heredity of the characters and mapping the related genes. There are serious reproductive barriers, mainly incrossability and hybrid sterility, in the interspecific hybridization of O. sativa with non-AA genome wild rice. These are the ＇bottleneck＇ for transferring elite genes from wild rice to O. sativa. Combining traditional crossing method with biotechnique is a reliable way to overcome the reproductive barriers and to improve the utilizing efficiency of non-AA genome wild rice.

Genetic Relationships in Advanced Generation Hybrids Derived from Crosses between Texas (<i>Poa arachnifera</i>) and Kentucky (<i>Poa pratensis</i>) Bluegrass Using ISSR Markers

Fertile, advanced generation hybrids derived from crosses between Texas (Poa arachnifera Torr.) and Kentucky (Poa pratensis L.) bluegrass have been selected. The hybrids are currently being evaluated for low-input turf potential. Since they are derived from hand-harvested seed from first-generation hybrids located in field nurseries their exact genetic origin is unknown. This experiment was conducted to determine if there are still genetic relationships among the advanced generation hybrids and some of the Texas and Kentucky parents in their pedigrees. Four Inter Simple Sequence Repeat (ISSR) primer combinations resolved on 6% nondenaturing polyacrylamide gels resulted in 179 polymorphic bands that were scored to create a genetic similarity matrix and dendrogram based on Jaccard’s coefficient. The clustering of the advanced generation hybrids was generally in agreement with what would be expected based on their pedigrees and indicated it was more likely to select a fertile hybrid from an advanced generation, rather than the F1 generation.

Genetic Variation and Heritability Estimates for Bunch Yield, Bunch Components and Vegetative Traits in Oil Palm Interspecific Hybrids

Progeny testing of oil palm interspecific hybrids was conducted in four trials in Kluang, Ulu Paka, Teluk Intan and Carey Island, Malaysia. The interspecific hybrids （O × P） were created according to the North Carolina Model I （NCM I） mating design, using Elaeis oleifera （0） and Elaeis guineensis var. pisifera （P） as the maternal and paternal parents, respectively. Differences among O, P and O-within-P were determined by the performance （bunch yield, components and vegetative traits） of the progenies. There were significant differences among P for fresh fruit bunch （FFB） yield, bunch number （BN） and average bunch weight （ABW） in Kluang, Ulu Paka and Teluk Intan, but not in Carey Island. FFB yield was generally higher on coastal soils （Teluk Intan and Carey Island） than inland soils （Kluang and Ulu Paka）. Heritability was calculated based on the intraclass correlation. Heritability estimates for these three yield components were variable, depending on the breeding material and environment in which the materials were tested. Fruit to bunch （F/B） and oil to bunch （O/B） of parthenocarpic fruits were important in determining the overall O/B of the interspecific hybrids. The O x P hybrids in Kluang showed the lowest height increment with only a mean of 14.0 cm/year, whereas in Ulu Paka and Teluk Intan, the values were higher at 24.0 cm/year and 25.0 cm/year, respectively. The study showed that the FFB yields ofoil palm interspecific hybrids performed better in coastal soils than inland soils.

Interspecific hybridization between Bursaphelenchus xylophilus and Bursaphelenchus mucronatus

Bursaphelenchus xylophilus is the pathogen that causes pine wilt disease, which has greatly damaged forests and ecosystems in countries of East Asia and Europe.Bursaphelenchus mucronatus is closely related to B. xylophilus in morphology and host plant specificity. A longrunning debate has existed regarding whether these two species can successfully produce hybrid offspring. In the present study, we performed in the laboratory, hybridization of two B. xylophilus nematode isolates from China and Japan and three B. mucronatus isolates from China, Japan and France. Nematode isolates of B. xylophilus were successfully crossed with B. mucronatus isolates, and the rate of hybridization was relatively high; however, some hybrid offspring died. Successful hybridization occurred between B. xylophilus and B. mucronatus isolates from China, and 22 generations of hybrids were produced. All F1 hybrids could be backcrossed with their parents and produce offspring. Variation in mucro length among the hybrid offspring and their parents was observed. The hybrid offspring and their parents were inoculated into 3-months-old black pine(Pinus tunbergii) seedlings. Weaker pathogenicity of hybrid offspring was observed compared with that of their parents, and significantly fewer offspring nematodes than parents were reisolated from pines. Therefore, the offspring of B. xylophilus and B. mucronatus may exist in the forest and could influence disease epidemics.

INTERSPECIFIC SOMATIC HYBRIDS IN ACTINIDIA

Protoplasts isolated from cotyledon-derived calli of Actinidia chinensis var. chinensis (2n = 2x=58) were fused by the PEG method with cotyledon-callus protoplasts of A. deliciosa var. deliciosa (2n = 6x = 174) or with mesophyll protoplasts of A. kolomikta (2n = 2x = 58), respectively. Randomly amplified polymorphic DNA (RAPD) markers and flow cytometry was used to confirm the occurrence of somatic hybrids. RAPD results with some primers surveyed indicated that one clone (A. chinensis + A. deliciosa) and four clones (A. chinensis + A. kolomikta) had RAPD banding patterns which combined the parental banding profiles. Ploidy levels of the (A. chinensis + A. deliciosa) clone were deduced as octoploid (2n = 8x), and the (A. chinensis + A. kolomikta) clones were tetraploid (2n = 4x), triploid (2n = 3x) or pentaploid (2n = 5x). The clones were confirmed as interspecific somatic hybrids in Actinidia.

Inheritance of the Anatomy--Morphological Structure of the Stalk by Interspecific Hybrids of the Glycine L.

The research focuses on the study of anatomical and morphological stalk structure ofsoya interspecific hybrids of the third generation （F3） between （Glycine max （L.） Merr.） and G. soja Sieb. et Zucc. in comparison with parent plant species. The parent plant species and interspecific hybrids were sowed and grew under similar conditions. The similarity of the anatomic structure of stalks of cultivated plants and Glycine soja （wild soya） proves the hypothesis the studied species have the same origin. However, the obtained results show the considerable degree of phylogenetic dissociation between the studied soya species. Interspecific hybrids inherit from G. soja the ability to high intensive growth. The G. soja use in practical selective breeding is of great interest.

Interspecific Hybridization Between Crassostrea angulata and C. ariakensis

lnterspecific hybridization can generate heterosis, which is proven to be a useful tool in selective breeding programs for oyster culture. Crassostrea angulata and C. ariakensis are two important economic shellfish species in China. We conducted 2 × 2 reciprocal crosses to determine whether these two species can cross-fertilize and their hybrids can hatch, survive and perform hetero- sis. Fertilization was found symmetrical without delay. The rate of fertilization success of C. angulata ♀× C. ariakensis ♂ was lower than that of C. ariakensis ♀ x C. angulata ♂, and the success rate of both hybridizations was lower than that of two intraspecific crosses each. During the planktonic period, survival rate of the progeny was lower in the hybrid crosses than in the intraspecific crosses. On day 360, mean shell height of the progeny of C. angulata ♀ x C. angulata ♂ was highest, which was followed by that of C. angulata ♀ × C. ariakensis ♂, C. ariakensis ♀x C. ariakensis ♂ and C. ariakensis ♀× C. angulata ♂ in a descending order. Mor- phology of adults produced by the hybrid crosses was similar to that of C. angulata. Both hybrids underwent normal gonad devel- opment and produced mature gametes in the mating season. This study provided new insights into the quantitative traits in interspeci- fic crosses of Crassostrea species, thus being of guidance value for selective breeding of oyster.

Cytogenetic Analysis of the Primary Amphidiploid Derived from Interspecific Hybridization in Cucumis and Its Selfed Progenies

Studies on the reproduction and cytogenetic characterization of a primary amphidiploid Cucumis species C. hytivus Chen and Kirkbride (2n = 4x = 38) indicated that a more comprehensive cytogenetic analysis of this species and its first selfed progeny would increase its potential utility in cucumber improvement. With tendrils used as source materials for mitotic analysis, chromosome numbers in all selfed progenies were 2n = 38, confirming chromosomal stability in this synthetic amphidiploid species. Detailed meiotic processes were described by comparing the primary and the selfed amphidiploids. Meiotic abnormalities, such as chromosome lagging, unequal separation, chromosome multi-polarization and polyads were observed frequently in all amphidiploid plants except for the selfed no.8, in which meiosis was arrested prior to metaphase Ⅰ. Generally, the frequency of multivalents was higher and the configurations were more complex in the selfed progenies, demonstrating a more extensive genetic exchange between cucumber and C. hystrix Chakr. Genome separation between cucumber and C. hystrix was observed through prophase Ⅰ to anaphase Ⅰ in both generations of the amphidiploids. Consequently, in addition to n = 19, a new gamete with n = 7 was produced, which was confirmed by the chromosome counts 2n = 14 in the backcrossing progenies from cucumber × amphidiploid mating. Fertility varied among the selfed amphidiploid plants. The selfed plant no.1 was found to have an improved fertility (e.g., pollen staining ability 40.8% and 25.6 seeds per fruit) and then was used as source germplasm in further introgression and gene exchange experiments.

Vacancy defect MoSeTe embedded in N and F co-doped carbon skeleton for high performance sodium ion batteries and hybrid capacitors

Sodium-ion batteries(SIBs) and hybrid capacitors(SIHCs) have garnered significant attention in energy storage due to their inherent advantages,including high energy density,cost-effectiveness,and enhanced safety.However,developing high-performance anode materials to improve sodium storage performa nce still remains a major challenge.Here,a facile one-pot method has been developed to fabricate a hybrid of MoSeTe nanosheets implanted within the N,F co-doped honeycomb carbon skeleton(MoSeTe/N,F@C).Experimental results demonstrate that the incorporation of large-sized Te atoms into MoSeTe nanosheets enlarges the layer spacing and creates abundant anion vacancies,which effectively facilitate the insertion/extraction of Na^(+) and provide numerous ion adsorption sites for rapid surface capacitive behavior.Additionally,the heteroatoms N,F co-doped honeycomb carbon skeleton with a highly conductive network can restrain the volume expansion and boost reaction kinetics within the electrode.As anticipated,the MoSeTe/N,F@C anode exhibits high reversible capacities along with exceptional cycle stability.When coupled with Na_(3)V_(2)(PO_(4))_(3)@C(NVPF@C) to form SIB full cells,the anode delivers a reversible specific capacity of 126 mA h g^(-1) after 100 cycles at 0.1 A g^(-1).Furthermore,when combined with AC to form SIHC full cells,the anode demonstrates excellent cycling stability with a reversible specific capacity of50 mA h g^(-1) keeping over 3700 cycles at 1.0 A g^(-1).In situ XRD,ex situ TEM characterization,and theoretical calculations(DFT) further confirm the reversibility of sodium storage in MoSeTe/N,F@C anode materials during electrochemical reactions,highlighting their potential for widespread practical application.This work provides new insights into the promising utilization of advanced transition metal dichalcogenides as anode materials for Na^(+)-based energy storage devices.

Key Role of Hybridization between Actinide 5<i>f</i>and Oxygen 2<i>p</i>Orbitals for Electronic Structure of Actinide Dioxides

In order to promote our understanding on electronic structure of actinide dioxides, we construct a tight-binding model composed of actinide 5f and oxygen 2p electrons, which is called f-p model. After the diagonalization of the f-p model, we compare the eigen-energies in the first Brillouin zone with the results of relativistic band-structure calculations. Here we emphasize a key role of f-p hybridization in order to understand the electronic structure of actinide dioxides. In particular, it is found that the position of energy levels of Г7 and Г8 states determined from crystalline electric field (CEF) potentials depends on the f-p hybridization. We investiagte the values of the Slater-Koster integrals for f-p hybridization, (fpσ) and (fpπ), which reproduce simultaneously the local CEF states and the band-structure calculation results. Then, we find that the absolute value of (fpπ) should be small in comparison with (fpσ) = 1 eV. The small value of |(fpπ)| is consistent with the condition to obtain the octupole ordering in the previous analysis of the f-p model.

Identification and cell wall analysis of interspecific hybrids between Oryza sativa and Oryza ridleyi

Oryza ridleyi is an allotetraploid wild species with the HHJJ genome, and Oryza sativa is a diploid cultivated rice that has the AA genome. Although the wide hybrid between the two species is difficult to obtain, we overcome this difficulty by young embryo rescue. An obvious heterosis was primarily found for the plant height, tillering ability, vegetative vigor, etc. However, the hybrid panicle and culm traits were found to resemble that of the wild rice parent, O. ridleyi, for the long awns, exoteric purple stigma, grain shattering, dispersed panicles, and culm mechanical strength. Genomic in situ hybridization （GISH） analysis was subsequently performed on the mitotic metaphase chromosome of the root tips, and we determined that the hybrid is an allotriploid with 36 chromosomes and its genomic constitution is AHJ. Chemical analyses conducted on the culm of O. sativa, O. ridleyi, and their interspecific hybrids showed that major changes occurred in the xylose, glucose, and arabinose concentrations, which are correlated with the specific hemicellulose polymer and cellulose components that are important in the primary cell walls of green plants. Meanwhile, the culm anatomical analyses indicated that additional large vascular bundles and an extra sclerenchyma cell layer were found in O. ridleyi. Additionally, further thickening of the secondary cell walls of the cortical fiber sclerenchyma cells and the phloem companion cells was discovered in O. ridleyi and in the interspecific hybrids. These results imply that there may be a potential link between culm mechanical strength and culm anatomical structure.

Interspecific Hybridization between Cucumis ssp. Plants and SRAP Analysis of Hybrids

To understand the inter-hybridization between Cucumis ssp.plants,we used 150 melon varieties as female parents to cross with Cucumis metuliferus and Cucumis anguria.Only melon accessions V2 and V129 set fruits,but seeds from fruits V2（V129）×C.metuliferus were abortive.A few of seeds from the bottom of fruit V2（V129）×C.anguria were fertile.Sequence-related amplified polymorphism（SRAP）molecular markers were used to analyze the progenies of inter-specific hybridization between C.anguria and melon V129.One pair primer（E14/M2）was found effective in amplification on male parent characteristic bands from the hybrids,suggesting that some DNA exchange had happened between C.anguria and melon V129.This study provided data for analyzing the mechanism of inter-hybridization between Cucumis plants.