Variations in chlorosis and potential usefulness of alloplasmic Brassica rapa with the cytoplasm of male sterile Brassica juncea

To select superior seed parents for vegetable hybrid seed production, we conducted interspecific crosses between male sterile Brassica juncea (2n = 36, AABB) and eight inbred lines of Brassica rapa (2n = 20, AA). Alloplasmic lines of B. rapa with the cytoplasm of B. juncea were developed from B. juncea × B. rapa hybrids by repeated backcrossing using B. rapa as the recurrent male parent until the BC₃ generation. Seed fertility, male sterility and chlorophyll content were investigated in these plants cultivated under four different temperature conditions (5, 10, 12 and 20°C). At 10°C, the alloplasmic lines of B. rapa with the cytoplasm of B. juncea were male sterile with partly chlorotic leaves. The alloplasmic B. rapa had lower chlorophyll a, chlorophyll b and carotenoid contents than those of the original B. rapa. The leaves recovered from chlorosis when the plants were cultivated at 20°C. An alloplasmic line of B. rapa (A6) is available as a seed parent for vegetable hybrid seed production and contributes seed fertility, slight chlorosis and stable male sterility.     

Stripe rust resistance gene Yr18 and its suppressor gene in Chinese wheat landraces

The slow‐rusting and mildewing gene Yr18/Lr34/Pm38/Sr57 confers partial, durable resistance to multiple fungal pathogens and has its origins in China. A number of diagnostic markers were developed for this gene based on the gene sequence, but these markers do not always predict the presence of the resistant phenotype as some wheat varieties with the gene are susceptible to stripe rust in China. We hypothesized that these varieties have a suppressor of Yr18. This study was undertaken to determine the presence of Yr18, the suppressor and/or another resistance gene in 144 Chinese wheat landraces using molecular markers and stripe rust field data. Forty‐three landraces were predicted to have Yr18 based on the presence of the markers, but had final disease severities higher than 70%, indicating that this gene may be under the influence of a suppressor. Four of these landraces, ‘Sichuanyonggang 2’, ‘Baikemai’, ‘Youmai’ and ‘Zhangsihuang’, were chosen for genetic studies. Crosses were made between the lines and ‘Avocet S’, with further crosses of Sichuanyonggang 2 ×  ‘Huixianhong’ and Sichuanyonggang 2 ×  ‘Chinese Spring’. The F₁ plants of Sichuanyonggang 2/Chinese Spring was susceptible indicating the presence of a dominant suppressor gene. The results of genetic analyses of F_2:3 and BC₁F₂ families derived from these crosses indicated the presence of Yr18, a Yr18 suppressor and another additive resistance gene. The Yr18 region in Sichuanyonggang 2 was sequenced to ensure that it contained the functional allele. This is the first report of a suppressor of Yr18/Lr34/Pm38/Sr57 gene with respect to stripe rust response.     

Phenotypic and seed structural comparison in hybrids of different Brassica rapa and B. oleracea

Brassica napus is an important oil species with short history and narrow genetic background. Interspecific hybrids from crosses between B. oleracea and different B. rapa were obtained. We found the hybrids with white petal resembling B. oleracea, the flavonoid and phenolic content decreased in hybrids, agreeing with the expressional changes of flavonoid biosynthesis genes. Seed coat of hybrids resembled diploid parents, or partly resembled to each parent with a clear outline. The palisade layer in hybrids was thicker than parents, with similar pigment accumulation as B. oleracea but more than B. rapa. Differentially sized protein bodies (PBs) were found in hybrids. The radical and inner cotyledon of all hybrids were identified with larger but less PBs than parents. The average size of PBs in outer cotyledon of resynthesized B. napus was also larger than parents, but the number of PBs was not significantly reduced. The phenotypic and seed structural variations after polyploidization of B. napus would be interesting for genetic broadening and breeding of rapeseed.     

Genetic Variation for Heat Tolerance During the Reproductive Phase in Brassica rapa

We investigated heat tolerance at the reproductive stage in six spring‐type B. rapa accessions and one B. juncea accession as a control. Plants were subjected to two temperature treatments for seven days in controlled environmental rooms, beginning one day before the first open flower on the main stem inflorescence. The high‐temperature treatment ranged from 25 °C to 35 °C during 16 h light and 25 °C during 8 h dark. The control temperature treatment was set at 23 °C during 16 h light and 15 °C during 8 h dark. Soil moisture was maintained at close to field capacity to avoid drought stress. Main stem buds that emerged during the treatment period were tagged, and pod and seed production was recorded at each reproductive node. Leaf temperature depression and leaf conductance increased in the high‐temperature treatment which indicated that plants were not drought stressed. A leafy vegetable type of B. rapa from Indonesia was the most tolerant to high temperature, as defined by its ability to set seed equally well in the control and high‐temperature treatments, followed by an oilseed type from Pakistan. Pollen viability remained above 87 % in all accessions and treatments. We conclude that bud number and length, and pod number produced under high temperatures, might provide a useful preliminary screen for high‐temperature tolerance and that B. rapa may be a valuable source of heat tolerance in canola (B. napus).     

Cytological observation of anther structure and genetic investigation of a new type of cytoplasmic male sterile 0A193‐CMS in Brassica rapa L.

Cytoplasmic male sterility (CMS), a maternally transmitted failure in pollen formation, is an effective pollination control system in hybrid rapeseed (Brassica napus) breeding. However, CMS is not widely used in the related oilseed species Brassica rapa. In the past years, several male sterile plants have been isolated from the B. rapa landrace ‘0A193’, collected in Shaanxi, China, in 2011. It is noteworthy that the fertility expression of 0A193‐CMS was affected by temperature. In contrast to pol CMS, fertility tests with 18 B. rapa and 9 B. napus accessions suggest that a different system of maintaining and restoring is responsible for the observed phenotype. Further on, genetic investigation evidenced that fertility of 0A193‐CMS is controlled by both cytoplasmic and one pair of nuclear recessive genes. Interestingly, plants of the 0A193‐CMS type possess a highly specific fragment of the mitochondrial gene orf222, a crucial regulator of male sterility in nap CMS. Our study broadens the CMS resources in B. rapa and provides a highly applicable alternative to pol CMS and ogu CMS for hybrid breeding production.     

Effect of Rht Alleles on the Tolerance of Wheat Grain Set to High Temperature and Drought Stress During Booting and Anthesis

Factorial pot experiments were conducted to compare the responses of GA‐sensitive and GA‐insensitive reduced height (Rht) alleles in wheat for susceptibility to heat and drought stress during booting and anthesis. Grain set (grains/spikelet) of near‐isogenic lines (NILs) was assessed following three day transfers to controlled environments imposing day temperatures (t) from 20 to 40 °C. Transfers were during booting and/or anthesis and pots maintained at field capacity (FC) or had water withheld. Logistic responses (y = c/1+e^{‐b(t ‐m)}) described declining grain set with increasing t, and t₅ was that fitted to give a 5 % reduction in grain set. Averaged over NIL, t₅ for anthesis at FC was 31.7 ± 0.47 °C (S.E.M., 26 d.f.). Drought at anthesis reduced t₅ by <2 °C. Maintaining FC at booting conferred considerable resistance to high temperatures (t₅ = 33.9 °C) but booting was particularly heat susceptible without water (t₅ = 26.5 °C). In one background (cv. Mercia), for NILs varying at the Rht‐D1 locus, there was progressive reduction in t₅ with dwarfing and reduced gibberellic acid (GA) sensitivity (Rht‐D1a, tall, 32.7 ± 0.72; Rht‐D1b, semi‐dwarf, 29.5 ± 0.85; Rht‐D1c, severe dwarf, 24.2 ± 0.72). This trend was not evident for the Rht‐B1 locus or for Rht‐D1b in an alternative background (Maris Widgeon). The GA‐sensitive severe dwarf Rht12 was more heat tolerant (t₅ = 29.4 ± 0.72) than the similarly statured GA‐insensitive Rht‐D1c. The GA‐sensitive, semidwarfing Rht8 conferred greater drought tolerance in one experiment. Despite the effects of Rht‐D1 alleles in Mercia on stress tolerance, the inconsistency of the effects over background and locus led to the conclusion that semidwarfing with GA‐insensitivity did not necessarily increase sensitivity to stress at booting and flowering. In comparison with effects of semidwarfing alleles, responses to heat stress are much more dramatically affected by water availability and the precise growth stage at which the stress is experienced by the plants.     

Mapping quantitative trait loci (QTLs) underlying seed vitamin E content in soybean with main,epistatic and QTL × environment effects

Soybean (Glycine max (L.) Merr.) seed contains small amounts of tocopherol, a non‐enzymatic antioxidant known as lipid‐soluble vitamin E (VE). Dietary VE contributes to a decreased risk of chronic diseases in humans and has several beneficial effects on resistance to stress in plants, and increasing VE content is an important breeding goal for increasing the nutritional value of soybean. In this study, quantitative trait loci (QTLs) underlying VE content with main, epistatic and QTL × environment effects were identified in a population of F_5 : 6 recombinant inbred lines from a cross between ‘Hefeng 25’ (a low‐VE cultivar) and ‘OAC Bayfield’ (a high‐VE cultivar). A total of 18 QTLs were detected that showed additive main effects (a) and/or additive × environment interaction effects (ae) in different environments. Moreover, 19 epistatic pairs of QTLs were found to be associated with α‐tocopherol (α‐Toc), γ‐tocopherol (γ‐Toc), δ‐tocopherol (δ‐Toc) and total VE (TE) contents. The QTLs identified in multienvironments could provide more information about QTL by environment interactions and could be useful for the marker‐assistant selection of soybean cultivars with high seed VE contents.     

Creating favourable morphological and yield variations for rapeseed by interspecific crosses between Brassica rapa and Brassica oleracea

Brassica napus is a most important oilseed grown worldwide with a limited genetic background, due to the short history of speciation, domestication and cultivation. To create novel germplasm for rapeseed breeding, we made interspecific crosses followed with chromosome doubling between B. rapa and B. oleracea to generate novel B. napus with favourable agronomic traits. The resynthesized (S0) hybrids were confirmed by SSR and cytogenetic analysis, and the fertility was increased from 32.7% in S0 generation to ~97.31% in S1 generation. The plant shapes of the progeny were dramatically improved compared to the diploid parents and B. napus cv. ‘Yangyou 6’, especially for the branch initiation height, branch number and pod number. The single‐plant yield was significantly improved in S1 progeny for the variations in branching sites and number. Significant improvement in plant shape and yield was observed on S2 generation compared to the local elite commercial open‐pollinated cultivar, which would be further fixed by intensive selection and pyramiding breeding. Such variation is of great value for breeding rapeseed with improved plant architecture and harvest index.     

Screening for resistance to black rot in a Spanish collection of Brassica rapa

Productivity and quality of crops of Brassica rapa L. in north‐western of Spain are highly affected by black rot, caused by the bacterium Xanthomonas campestris pv. campestris (Pammel) Dowson (Xcc). Several races of Xcc have been described in this area, being the race 6 the most frequent in B. rapa crops and races 1 and 4 the most frequent in B. oleracea crops. The control of the disease can be aided by the employment of resistant varieties. The aim of this work was to find sources of resistance to Xcc in a collection of open‐pollinated varieties of B. rapa from north‐western Spain. Resistance was evaluated in 191 landraces. Partial resistance to races 6, 1 and 4 and complete resistance to race 4 were identified in several landraces. Several accessions exhibited partial resistance to the three races. Sources of resistance were identified in landraces of different crops of the species (turnips, turnip greens and turnip tops). These landraces could be grown after selection for resistance or they can be donors of resistance genes in breeding programmes.     

Rapid generation cycling of an F2 population derived from a cross between Lens culinaris Medik. and Lens ervoides (Brign.) Grande after aphanomyces root rot selection

Cultivated lentil (Lens culinaris Medik.) is susceptible to aphanomyces root rot (ARR), whereas partial resistance is present in wild lentil including Lens ervoides (Brign.) Grande. Approximately six generations of selfing are required to fix a desired trait in a population, which usually requires 2 years in a breeding programme, so the primary objective was to develop a rapid generation cycling (RGC) technique that achieves this goal in 1 year. Rapid generation cycling was then tested on an F₂ population (LR‐59) derived from a L. culinaris × L. ervoides cross in combination with a reliable ARR screening technique, which generates a wide range of disease severities conducive to selection. Phenotyping of an F₂ population of more than 1,200 plants resulted in scores ranging from 2.4 to 4.0 on a scale from zero to five. Plants with scores lower than 4.0 were selected for advancement for five generations using a modified single‐seed descent method, optimum growing conditions, 20‐hr photoperiod and harvest of immature seeds. Seeds were germinated in a 100 μM gibberellin solution. Average generation length after phenotyping was 56 days resulting in five generations within approximately 300 days. Using a modified inoculation protocol, ARR phenotyping of the F₇ population resulted in scores ranging from 1.4 to 4.0. This inexpensive, nonsterile speed breeding protocol saves 1 year in the development of lentil varieties with improved ARR resistance.     

Inheritance of resistance to the peach root‐knot nematode (Meloidogyne floridensis) in interspecific crosses between peach (Prunus persica) and its wild relative (Prunus kansuensis)

The peach root‐knot nematode, Meloidogyne floridensis (MF), infects majority of available nematode‐resistant peach rootstocks which are mostly derived from peach (Prunus persica) and Chinese wild peach (P. davidiana). Interspecific hybridization of peach with its wild relative, Kansu peach (P. kansuensis), offers potential for broadening the resistance spectrum in standard peach rootstocks. We investigated the inheritance of resistance to MF in segregating populations of peach (‘Okinawa’ or ‘Flordaguard’) × P. kansuensis. A total of 379 individuals from 13 F₂ and BC₁F₁ families were challenged with a pathogenic MF isolate “MFGnv14” and were classified as resistant (R) or susceptible (S) based on root galling intensity. Segregation analyses in F₂ progeny revealed the involvement of a major locus with a dominant or recessive allele determining resistance in progeny segregating 3R:1S and 1R:3S, respectively. Testcrosses with a homozygous‐susceptible peach genotype (‘Flordaguard’ or ‘UFSharp’) confirmed P. kansuensis as a source of new resistance and the heterozygous allelic status of P. kansuensis at the locus conferring resistance to MF. We propose a single‐locus dominant/recessive model for the inheritance of resistance.     

Mechanisms and diversity of resistance to sorghum shoot fly,Atherigona soccata

Sorghum shoot fly, Atherigona soccata, is one of the important pests of postrainy season sorghums. Of the 90 sorghum genotypes evaluated for resistance to this pest, RHRB 12, ICSV 713, 25026, 93046 and 25027, IS 33844‐5, Giddi Maldandi and RVRT 3 exhibited resistance in postrainy season, while ICSB 463, Phule Anuradha, RHRB 19, Parbhani Moti, ICSV 705, PS 35805, IS 5480, 5622, 17726, 18368 and 34722, RVRT 1, ICSR 93031 and Dagidi Solapur showed resistance in rainy season, suggesting season‐specific expression of resistance to A. soccata. ICSB 461, ICSB 463, Phule Yasodha, M 35‐1, ICSV 700, 711, 25010, 25019 and 93089, IS 18662, Phule Vasudha, IS 18551 and 33844‐5 and Barsizoot had fewer deadhearts than plants with eggs across seasons, suggesting antibiosis as one of the resistance mechanism. Five genotypes exhibited resistance with high grain yield across seasons. Correlation, path and stepwise regression analyses indicated that leaf glossiness, seedling vigour, trichome density, oviposition and leaf sheath pigmentation were associated with the expression of resistance/susceptibility to shoot fly, and these can be used as marker traits to select and develop shoot fly‐resistant sorghums.     

Production and characterization of Raphanus sativus-Brassica rapa monosomic chromosome addition lines

Breeding of Raphanus sativus‐Brassica rapa monosomic chromosome addition lines (MALs, 2n = 19) was carried out by backcrossing the synthesized amphidiploid line, Raphanobrassica (R. sativus×B. rapa, 2n = 38, RRAA, line RA89) with R. sativus cv. ‘Shogoin’ (2n = 18, RR). In the first cross of Raphanobrassica× radish, four sesquidiploidal BC₁ plants (2n = 28, RRA, RA89‐36‐1, RA89‐31‐1, RA89‐31‐2, RA89‐31‐3) were successfully developed. In these plants, the chromosome configurations of 9II + 10I and 10II + 8I were observed frequently at first metaphase (MI) of meiosis in pollen mother cells (PMCs). The RA 89‐36‐1 plant produced many seeds in the reciprocal backcrosses with radish. About 50% of the BC₂ plants obtained from the cross of RA89‐36‐1 plant × radish were 2n = 19 plants, followed by 2n = 18 plants (24%) and 2n = 20 plants (19%). In the reciprocal cross, 2n = 19 plants were also developed at the rate of 40%. From analysis of specific morphological traits, 2n = 19 plants were classified into eight types (a‐h). When 25 selected primers were used in polyacrylamide gel electrophoresis, random amplified polymorphic DNA (RAPD) markers derived from B. rapa for each type of MAL were detected in numbers between three for e‐type and 16 for b‐type. RAPD markers specific for each type alone were from one (OPE 05‐344) for h‐type to nine for b‐type. In the g‐type, no marker specific to this type alone was observed. However, 19 bands were common between at least two types. These MAL plants exhibited predominantly the chromosome configuration of 9II + 1I at MI of PMCs, pollen and seed fertility being the same level as the radish cv. ‘Shogoin’. From the morphological traits and DNA markers, eight different MAL types among 10 expected were identified.     

Genotypic Variation for Tolerance to Transient Drought During the Reproductive Phase of Brassica rapa

Brassica rapa L. is a genetically diverse parent species of the allotetraploid species, oilseed rape (B. napus) and a potential source of drought tolerance for B. napus. We examined the effect of a 13‐day drought stress period during the early reproductive phase, relative to a well‐watered (WW) control, on subsequent growth and development in nine accessions of B. rapa and one accession of Brassica juncea selected for their wide morphological and genetic diversity. We measured leaf water potential, stomatal conductance, water use, and leaf and bud temperatures during the stress period and aboveground dry weight of total biomass at maturity. Dry weight of seeds and reproductive tissue were not useful measures of drought tolerance due to self‐incompatibility in B. rapa. The relative total biomass (used as the measure of drought tolerance in this study) of the 10 accessions exposed to drought stress ranged from 47 % to 117 % of the WW treatment and was negatively correlated with leaf‐to‐air and bud‐to‐air temperature difference when averaged across the 13‐day stress period. Two wild‐type (B. rapa ssp. sylvestris) accessions had higher relative total and non‐reproductive biomass at maturity and cooler leaves and buds than other types. We conclude that considerable genotypic variation for drought tolerance exists in B. rapa and cooler leaves and buds during a transient drought stress in the early reproductive phase may be a useful screening tool for drought tolerance.     

Development of molecular markers linked to the Leptosphaeria maculans resistance gene Rlm6 and inheritance of SCAR and CAPS markers in Brassica napus × Brassica juncea interspecific hybrids

Leptosphaeria maculans causes blackleg disease on Brassica napus, an economically important oilseed crop. Brassica juncea has high resistance to blackleg and is a source for the development of resistant B. napus varieties. To transfer the Rlm6 resistance gene from B. juncea into B. napus, an interspecific cross between B. napus “Topas DH16516” and B. juncea “Forge” was produced, followed by the development of F₂ and F₃ generations. Sequence characterized amplified region (SCAR) and cleaved amplified polymorphic sequence (CAPS) markers linked to the L. maculans resistance gene Rlm6 were developed. Segregation of SCAR and CAPS markers linked to Rlm6 were confirmed by genotyping of F₂ and F₃ progeny. Segregation of CAPS markers and phenotypes for blackleg disease severity in F₂ plants had a Mendelian ratio of 3:1 in resistant vs. susceptible plants, respectively, supporting the assumption that genetic control of resistance was by a single dominant gene. The molecular markers developed in this study, which show linkage with the L. maculans resistance gene Rlm6, would facilitate marker‐assisted backcross breeding in a variety development programme.     

Identification of an ice recrystallisation inhibition gene family in winter‐hardy wheat and its evolutionary relationship to other members of the Triticeae

Ice recrystallisation inhibition (IRI) proteins are important for plants that exist in cold environments. Six novel genes encoding IRI proteins were isolated from winter‐hardy wheat (Tricticum aestivum) cultivar (Mironovskaya808). Conserved domain analysis confirmed that all the predicted proteins contained the basic characteristics of IRI proteins, including leucine‐rich repeats (LRR) and IRI‐domains (NxVx_1‐2G). In addition, these proteins possessed an N‐terminal signal peptide and some conserved cysteines. Despite the high conservation of the amino acid sequences, the characteristic regions still exist among these IRI‐domains of IRI sequences. All IRI sequences in this study are highly homologous with phytosulfokine receptors (PSR). Quantitative real‐time PCR (qPCR) analysis indicated that each member of the IRI gene family was induced after exposure to low temperature. Heterologous expression of IRI proteins enhanced freezing tolerance in host cells. A total of 28 IRI genes were isolated from Triticeae, and the evolutionary relationship of the IRI gene family in Triticeae and related species showed high interspecific specificity. The IRI genes from Aegilops, which is one of the progenitors of allohexaploid wheat, shared the closest relationship with those from Triticum. With high interspecific specificity in Triticeae, however, there was no specificity among different species of Triticum. Our results suggest that IRI genes have potential value to improve cold resistance of crops.     

Identification and marker‐assisted introgression of QTL conferring resistance to bacterial leaf blight in cowpea (Vigna unguiculata (L.) Walp.)

Bacterial leaf blight (BLB), caused by Xanthomonas axonopodis pv. vignicola (Xav), is widespread in major cowpea [Vigna unguiculata (L.) Walp.] growing regions of the world. Considering the resource poor nature of cowpea farmers, development and introduction of cultivars resistant to the disease is the best option. Identification of DNA markers and marker‐assisted selection will increase precision of breeding for resistance to diseases like bacterial leaf blight. Hence, an attempt was made to detect QTL for resistance to BLB using 194 F_2 : 3 progeny derived from the cross ‘C‐152’ (susceptible parent) × ‘V‐16’ (resistant parent). These progeny were screened for resistance to bacterial blight by the leaf inoculation method. Platykurtic distribution of per cent disease index scores indicated quantitative inheritance of resistance to bacterial leaf blight. A genetic map with 96 markers (79 SSR and 17 CISP) constructed from the 194 F₂ individuals was used to perform QTL analysis. Out of three major QTL identified, one was on LG 8 (qtlblb‐1) and two on LG 11 (qtlblb‐2 and qtlblb‐3). The PCR product generated by the primer VuMt337 encoded for RIN2‐like mRNA that positively regulate RPM1‐ and RPS2‐dependent hypersensitive response. The QTL qtlblb‐1 explained 30.58% phenotypic variation followed by qtlblb‐2 and qtlblb‐3 with 10.77% and 10.63%, respectively. The major QTL region on LG 8 was introgressed from cultivar V‐16 into the bacterial leaf blight susceptible variety C‐152 through marker‐assisted backcrossing (MABC).     

Production of a desirable Brassica oleracea CMS line using an alloplasmic B. rapa CMS line carrying Diplotaxis erucoides cytoplasm as a bridge plant

In Brassica oleracea, production of F₁ hybrid seeds mainly makes use of the improved Ogura cytoplasmic male sterile (CMS) line. However, reliance on one particular line is a risk, and it would be advantageous to develop other CMS lines. In this study, we transferred Diplotaxis erucoides cytoplasm to B. oleracea cultivars using an alloplasmic B. rapaCMS line as a bridge plant to avoid incompatibility between donor and recipient plants. The new B. oleraceaCMS lines, which were derived by four generations of backcrossing, had small rudimentary anthers with no pollen grain and showed complete male sterility. There was no functional defect in other floral organs, and the ability to receive normal pollen did not appear to be impaired. Moreover, the B. oleraceaCMS lines carrying D. erucoides cytoplasm had larger leaf areas and a normal plastochron. As a consequence, the B. oleraceaCMS lines carrying D. erucoides cytoplasm have the potential to be valuable alternatives for use in commercial B. oleracea hybrid seed production.     

Validation of a male‐linked gene locus (OGI) for sex identification in persimmon (Diospyros kaki Thunb.) and its application in F1 progeny

It is crucial to develop a rapid technique for identifying sexuality in the seedling stage of persimmon (Diospyros kaki Thunb.), and the elimination of male progeny has been regarded as an important strategy for enhancing breeding efficiency. In this study, phenotype characterization and genotyping of the male‐linked OGI marker were carried out using 205 accessions, including persimmon cultivars, F₁ progeny and nine related Diospyros species. All persimmon cultivars displayed consistent results regarding OGI amplification and sex phenotype. A total of 143 F₁ progeny were derived from 11 crosses, among which 95 individuals had flowered. In the flowering full‐sib families, the amplification of the OGI marker in agreement with the sex phenotype was obtained in 85 plants (89.5%). The segregation of OGI in ‘Huashi 1’ × ‘Luotian Tianshi’ and ‘Huashi 1’ × Male 3 F₁ populations fit a 1 : 1 ratio. Furthermore, high OGI transferability was observed in nine related species. Overall, the results indicated that the OGI locus could be used to distinguish male from female persimmon plants at an early stage.