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Estimation of mean performance and heterosis in cytoplasmic male sterility (CMS) based hybrids in cabbage

Abstract

The successful transfer of R-cytoplasm (Ogura)-induced cytoplasmic male sterility has aroused great interest in the development of hybrids in cabbage. An experiment was conducted from 2018 to 2020 to study the per se performance and estimate heterosis in cabbage (Brassica oleracea L. var. capitata L.), This crop is highly cross-pollinated; thus, exploitation of heterosis is an additional advantage. The experimental material consisted of an F1 population of 18 crosses, resulting from three lines (CMS) crossed with six testers in a Line × Tester design, plus a standard check (Pusa Cabbage Hybrid-1). The per se performance and estimates of heterosis of the cross combinations L1 × T2, L2 × T2, L2 × T5, L3 × T2 and L3 × T5 were high for various horticultural parameters. Hence, after multilocation testing, these parents and crosses can be released to substitute existing cabbage varieties/hybrids.

Keywords:
R-cytoplasm; cabbage; cytoplasmic male sterility; heterosis; line × tester design

INTRODUCTION

Cabbage (Brassica oleracea L. var. capitata L.), with a diploid chromosome number (2n = 2x = 18), is one of the most important cole crops grown in the world. Modern cabbage cultivars were derived from wild, non-heading Brassica (Brassica oleraceae L. var. oleraceae L.) by mutation, human selection and adaptation. The crop is grown for its leafy heads, which are commonly consumed as salad, cole slaw, or boiled, pickled or dehydrated vegetable. This vegetable contains the indole-3-carbinol compound, which serves as protection against bowel cancer (Thamburaj and Singh 2001Thamburaj S, Singh N2001 Textbook of vegetables, tuber crops and spices. ICAR Publication, New Delhi, 99p).

Initially, only the sporophytic self-incompatibility system was used for commercial hybrid seed production. In the last decade, however, after the successful transfer of the R-cytoplasm (Ogura)-induced CMS system, the possibility of developing hybrids using cytoplasmic male sterility (CMS) has aroused great interest. Ogura CMS was inserted into cabbage by distant hybridization between radish and cabbage combined with embryo rescue, and OguraR1 CMS cabbage was created (Bannerot et al. 1974Bannerot H, Boulidard L, Cauderon Y, Tempe J1974 Transfer of cytoplasmic male sterility from Raphanus sativus to Brassica oleracea. Proceedings of Eucarpia meeting - Cruciferae 25:52-54). However, because Ogura CMS cabbage lines have a radish cytoplasm, problems with nuclear-cytoplasmic incompatibility, chlorosis at low temperature (15 ºC), nectary dysplasia and poor seed set occur frequently. OguraR2 CMS materials were successfully established by protoplast fusion, by which the radish chloroplast was replaced by that of cauliflower in the OguraR1 CMS system (Walters et al. 1992Walters TW, Mutschler MA, Earle ED1992 Protoplast fusion-derived Ogura male sterile cauliflower with cold tolerance. Plant Cell Reports 10:624-628). With Ogura CMSR3 material as the sterile source, several Ogura CMS cabbage lines with stable sterility and normal flowering and seed set have been bred and widely used in the development and production of cabbage hybrids (Ren et al. 2022Ren W, Si J, Chen L, Fang Z, Zhuang M, Lv H, Wang Y, Ji J, Yu H, Zhang Y2022 Mechanism and utilization of ogura cytoplasmic male sterility in cruciferae crops. International Journal of Molecular Sciences 23:9099).

Compared to the self-incompatible lines, cytoplasmic male sterile lines are more stable and could increase the cultivar purity of cabbage hybrids by 5-7% (Ding and Jian 2008Ding Y, Jian Y2008 Cabbage cytoplasmic male sterile line breeding and its application in hybrid seed production. Acta Horticulturae 71:89-95). This is a promising alternative approach that could overcome the problem of selfs in hybrid seed due to the breakdown of self-incompatibility and vigor decline in S-allele lines, as a result of repeated selfing and sib-mating.

The rate of conversion from open-pollinated to hybrid varieties and seed replacement ratio are very high in cabbage. So far, of all coordinated research projects on vegetable crops in India, only the regional research station ICAR-IARI, in Katrain, Kullu Valley, Himachal Pradesh, India, developed one CMS-based hybrid KTCH-1 (Pusa Cabbage Hybrid-1). In, view of the above facts, this study has been undertaken to assess the per se performance and heterosis for various horticultural traits of F1 population of 18 crosses, resulting from three CMS lines crossed with six testers in a Line × Tester design, plus a standard check.

MATERIAL AND METHODS

The experiment was carried out on an experimental farm of the Vegetable Science Department, Dr YS Parmar University of Horticulture and Forestry, Nauni, Solan, Himachal Pradesh, India, from 2018 to 2020. The parental lines were selected for their variability in various horticultural characteristics and practical applicability in heterosis breeding. The healthy seedlings of three lines viz., L1, L2 and L3, and six testers viz., T1, T2, T3, T4, T5 and T6, were transplanted to the main field on September 25, 2018. Details of the studied plant material are listed in Table 1. At proper head maturity in the last week of December 2018, the compact heads were incised crosswise, to facilitate bolting and flowering stalk emergence from the center of the heads. Crosses between the lines (three CMS lines) and testers (six) were initiated in February, 2019. Eighteen F1 hybrid combinations were developed by standard procedures of crossing and bud pollination, according to the Line × Tester mating design (Kempthorne 1957Kempthorne D1957 An introduction to genetic statistics. John Wiley & Sons, New York, p. 468-471). The matured siliquae of hybrids and parental lines were harvested separately from May to June, 2019. The seeds were dried and stored in a cool place for sowing in the following year.

Table 1
Identification of cabbage genotypes used in the study

On September 5, 2019, the seeds of the F1 hybrids (18), parents (three lines and six testers) along with a standard check (Pusa Cabbage Hybrid-1) were sown in a nursery. Healthy seedlings were transplanted to the main field on September 26, 2019, in a randomized complete block design (RCBD), with three replications. The following horticultural parameters were recorded: days to 50% marketable maturity, number of non-wrapper leaves, plant spread (cm), head compactness (g cm-3), head shape index, gross head weight (g), net head weight (g), stalk length (cm), harvest index (%), equatorial head diameter (cm), polar head diameter (cm), core size (cm), total soluble solids (˚B), ascorbic acid (mg) and yield per plot (kg) and per hectare (q).

RESULTS AND DISCUSSION

The mean performances of nine parents, 18 crosses and the standard check are shown in Tables 2-4. The mean squares due to parents and hybrids were found to be significant for all traits, which indicates the existence of sufficient variation among parents and hybrids, a prerequisite to improve traits through breeding. The mean and range of the parents were greater than of the hybrids for days to 50% maturity, number of wrapper leaves, plant spread (Table 2) and core size (Table 3), as shown by the desirable negative heterosis.

Table 2
Mean performance of parents, hybrids and standard check for the traits days to 50% marketable maturity, number of non-wrapper leaves, plant spread, head compactness, head shape index and gross head weight
Table 3
Mean performance of parents, hybrids and standard check for net head weight, stalk length, harvest index, equatorial head diameter, polar head diameter and core size

The mean and range of the F1 hybrids were found to be greater than those of the parents for head compactness, head shape index, gross head weight (Table 2), net head weight, polar diameter (Table 3), total soluble solids, ascorbic acid and yield per plot (Table 4). This can also be explained by the positive heterosis, which is desirable for these traits. The parental mean and range were lower than the mean and range of hybrids for stalk length (for which negative heterosis is desirable) and greater than the mean and range of hybrids for harvest index, equatorial diameter (Table 3) and total soluble solids (Table 4), which indicated superiority of the parents over hybrids.

Table 4
Mean performance of parents, hybrids and standard check for total soluble solids, ascorbic acid, yield per plot and yield per hectare

The significant superiority of 11 and 16 hybrids over the better parent and standard check, respectively, for days to 50% marketable maturity (for which negative heterosis is desirable) indicated dominant earliness and prospects to breed short duration hybrids to drive up the market price (Table 5). Significant desirable negative heterosis estimates for heterobeltiosis and standard heterosis were found for 11 cross combinations. Similar results were described by Parkash et al. (2015Parkash C, Dey SS, Bhatia R, Dhiman MR2015 Indigenously developed SI and CMS lines in hybrid breeding of cabbage. Indian Journal of Horticulture 72:212-217). Comparatively, a smaller number of non-wrapper leaves is considered desirable in cabbage which generally has shorter frame leaves. Significant desirable negative heterosis was recorded for heterobeltiosis and standard heterosis, respectively, in nine and one crosses. For one cross combination, significant desirable negative heterosis was estimated for heterobeltiosis and standard heterosis. These findings are in line with those of Thakur and Vidyasagar (2016aThakur H, Vidyasagar Vidyasagar2016a Estimates of genetic variability, heritability and genetic advances for yield and horticultural traits in cabbage (Brassica oleracea L. var. capitata L.). Journal of Environment and Bio-Sciences 30:155-157) and Kumar et al. (2019Kumar N, Chadha S, Kanwar S2019 CMS and SI based heterosis for yield and related traits in low chill cabbage under mid hills condition of Himachal Pradesh. Indian Journal of Horticulture 76:663-671).

Table 5
Heterobeltiosis and standard heterosis for days to 50% maturity, number of non-wrapper leaves, plant spread, head compactness and head shape index

Cabbage lines with reduced plant spread are desirable, since a greater number of plants per square meter can be accommodated, resulting in higher yields. Significant desirable negative heterosis for heterobeltiosis was recorded in five crosses. None of the cross combinations showed significant desirable negative heterosis estimates over standard check (Table 5). These findings are similar to those of Kumar et al. (2019Kumar N, Chadha S, Kanwar S2019 CMS and SI based heterosis for yield and related traits in low chill cabbage under mid hills condition of Himachal Pradesh. Indian Journal of Horticulture 76:663-671). High head compactness is desirable since a more compact head will have less volume and more weight per unit area. Of the 18 F1 hybrids, heterosis over the better parent and standard check, respectively, was significantly positive for 5 and 16 crosses. Five crosses showed significant desirable positive heterosis estimates for heterobeltiosis and standard heterosis, respectively. These findings were similarly reported by Singh et al. (2009Singh BK, Sharma SR, Singh B2009 Heterosis for antioxidants and horticultural traits in single cross hybrids of cabbage (Brassica oleracea L. var. capitata L.). Indian Journal of Agricultural Sciences 79:703-708) and Kumar et al. (2019). For head shape index, positive heterosis is desirable. Among the 18 F1 hybrids, heterosis was significant and positive for four and three crosses, respectively, over the better parent and standard check. Three crosses showed significant desirable positive heterosis estimates for heterobeltiosis and economic heterosis. Similar results were also obtained by Kumar et al. (2019) and Thakur and Vidyasagar (2016bThakur H, Vidyasagar Vidyasagar2016b Combining ability, gene action and heterosis studies involving SI and CMS lines and testers in cabbage. Green Farming 7:580-585).

High gross head weight is the key trait that contributes to high yields. Significant desirable negative heterosis was recorded for 2 and 14 crosses, respectively, for heterobeltiosis and standard heterosis (Table 6). For two crosses, the (desirable) positive heterosis values, respectively, were significant for heterobeltiosis and standard heterosis. Significant positive heterosis (desirable) for gross head weight in cabbage was also reported by Singh et al. (2009Singh BK, Sharma SR, Singh B2009 Heterosis for antioxidants and horticultural traits in single cross hybrids of cabbage (Brassica oleracea L. var. capitata L.). Indian Journal of Agricultural Sciences 79:703-708) and Parkash et al. (2015Parkash C, Dey SS, Bhatia R, Dhiman MR2015 Indigenously developed SI and CMS lines in hybrid breeding of cabbage. Indian Journal of Horticulture 72:212-217).

Table 6
Heterobeltiosis and standard heterosis for gross head weight, net head weight, stalk length, harvest index and equatorial head diameter

Net head weight is an important yield-related horticultural trait. Among the 18 F1 hybrids, for eight and nine crosses, respectively, heterosis was significant and positive over the better parent and standard check. For six crosses, heterosis for heterobeltiosis and standard heterosis was significant and positive (desirable). Similar results were obtained by Kumar et al. (2019Kumar N, Chadha S, Kanwar S2019 CMS and SI based heterosis for yield and related traits in low chill cabbage under mid hills condition of Himachal Pradesh. Indian Journal of Horticulture 76:663-671). For stalk length, negative heterosis is desirable. Significant desirable negative heterosis was recorded in one and 15 crosses for heterobeltiosis and standard heterosis (Table 6). One cross combination showed significant desirable negative heterosis estimates for heterobeltiosis and standard heterosis. Significant negative heterosis (desirable) for stalk length in cabbage was also reported by Thakur and Vidyasagar (2016bThakur H, Vidyasagar Vidyasagar2016b Combining ability, gene action and heterosis studies involving SI and CMS lines and testers in cabbage. Green Farming 7:580-585). Among the 18 F1 hybrids, two and three crosses showed desirable heterosis over the better parent and standard check, respectively, for harvest index (Table 5). Significant desirable positive heterosis values for heterobeltiosis and standard heterosis were not found for any of the crosses. These results are similar to those of Parkash et al. (2015Parkash C, Dey SS, Bhatia R, Dhiman MR2015 Indigenously developed SI and CMS lines in hybrid breeding of cabbage. Indian Journal of Horticulture 72:212-217).

For equatorial and polar head diameter, positive heterosis is desirable. Significant positive heterosis over the better parent and standard check, respectively, was recorded for equatorial diameter in one and two crosses (Table 6). One cross combination showed significant and positive heterosis (desirable) for heterobeltiosis and standard heterosis. Significant positive heterosis over the better parent and standard check was recorded in two and two crosses, respectively, for polar diameter (Table 7). For two crosses, significant positive heterosis (desirable) was estimated for heterobeltiosis and standard heterosis. These findings confirm those of Kibar et al. (2015Kibar B, Karaagac O, Kar H2015 Heterosis for yield contributing head traits in cabbage (Brassica oleracea L. var. capitata L.). Scientia Investigation Agraria 42:205-216) and Thakur and Vidyasagar (2016bThakur H, Vidyasagar Vidyasagar2016b Combining ability, gene action and heterosis studies involving SI and CMS lines and testers in cabbage. Green Farming 7:580-585).

Table 7
Heterobeltiosis and standard heterosis for polar head diameter, core size, total soluble solids, ascorbic acid and yield per plot

A relatively smaller core size is desirable as it highly corrected with compact head. The significant superiority for core size of three and six hybrids over the better parent and standard check, respectively, indicate the dominance of compact head for better shelf life of cabbage (Table 7). For three crosses, significant negative heterosis (desirable) was estimated for heterobeltiosis and standard heterosis. Three crosses had significant negative heterosis values (desirable) for heterobeltiosis and standard heterosis. Significant negative heterosis (desirable) for core size in cabbage was also reported by Parkash et al. (2015Parkash C, Dey SS, Bhatia R, Dhiman MR2015 Indigenously developed SI and CMS lines in hybrid breeding of cabbage. Indian Journal of Horticulture 72:212-217), Thakur and Vidyasagar (2016bThakur H, Vidyasagar Vidyasagar2016b Combining ability, gene action and heterosis studies involving SI and CMS lines and testers in cabbage. Green Farming 7:580-585) and Kumar et al. (2019Kumar N, Chadha S, Kanwar S2019 CMS and SI based heterosis for yield and related traits in low chill cabbage under mid hills condition of Himachal Pradesh. Indian Journal of Horticulture 76:663-671). For total soluble solids, positive heterosis is desirable. Significant positive heterosis over the better parent and standard check, respectively, was recorded in one and eighteen crosses. In one cross combination, significant desirable positive heterosis for this trait was estimated for heterobeltiosis and standard heterosis. These findings agree with those of Kumar et al. (2019Kumar N, Chadha S, Kanwar S2019 CMS and SI based heterosis for yield and related traits in low chill cabbage under mid hills condition of Himachal Pradesh. Indian Journal of Horticulture 76:663-671). Significant positive heterosis (desirable) for ascorbic acid of six and five hybrids over the better parent and standard check, respectively, was recorded. Significant positive heterosis (desirable) was estimated for five crosses for heterobeltiosis and standard heterosis. Similar trends were reported by Parkash et al. (2015Parkash C, Dey SS, Bhatia R, Dhiman MR2015 Indigenously developed SI and CMS lines in hybrid breeding of cabbage. Indian Journal of Horticulture 72:212-217). Yield is the prime and complex trait. For yield per plot, positive heterosis is desirable. Significant positive heterosis over the better parent and standard check was recorded in eight and nine crosses, respectively, which enlightening that there is enough scope to increase the productivity. Significant positive heterosis (desirable) for heterobeltiosis and standard heterosis was found for six crosses, in agreement with findings of Thakur and Vidyasagar (2016b).

CONCLUSION

The per se performance and heterosis estimates (better parent and standard check) indicated that the crosses L1 × T2, L2 × T5, L3 × T5, L3 × T2 and L2 × T2 performed well for various horticulture parameters (Supplementary material I). Thus, the results of this study might be relevant and useful for developing hybrids with good head compactness, harvest index, equatorial and polar diameter, net head weight, total soluble solids and ascorbic acid, to possibly enhance the plant stand per square meter and biotic and abiotic stress tolerance, without losing vigour advantage of the yield. Hence, after multilocation testing, these parents and crosses can be released to replace existing cabbage varieties/hybrids.

ACKNOWLEDGMENTS

The authors are indebted to the Department of Vegetable Science, Dr. YS Parmar University of Horticulture and Forestry, Nauni, Solan, Himachal Pradesh, India, for funding the research work. Supplementary material I can be provided upon request by the corresponding author.

REFERENCES

  • Bannerot H, Boulidard L, Cauderon Y, Tempe J1974 Transfer of cytoplasmic male sterility from Raphanus sativus to Brassica oleracea. Proceedings of Eucarpia meeting - Cruciferae 25:52-54
  • Ding Y, Jian Y2008 Cabbage cytoplasmic male sterile line breeding and its application in hybrid seed production. Acta Horticulturae 71:89-95
  • Kempthorne D1957 An introduction to genetic statistics. John Wiley & Sons, New York, p. 468-471
  • Kibar B, Karaagac O, Kar H2015 Heterosis for yield contributing head traits in cabbage (Brassica oleracea L. var. capitata L.). Scientia Investigation Agraria 42:205-216
  • Kumar N, Chadha S, Kanwar S2019 CMS and SI based heterosis for yield and related traits in low chill cabbage under mid hills condition of Himachal Pradesh. Indian Journal of Horticulture 76:663-671
  • Parkash C, Dey SS, Bhatia R, Dhiman MR2015 Indigenously developed SI and CMS lines in hybrid breeding of cabbage. Indian Journal of Horticulture 72:212-217
  • Ren W, Si J, Chen L, Fang Z, Zhuang M, Lv H, Wang Y, Ji J, Yu H, Zhang Y2022 Mechanism and utilization of ogura cytoplasmic male sterility in cruciferae crops. International Journal of Molecular Sciences 23:9099
  • Singh BK, Sharma SR, Singh B2009 Heterosis for antioxidants and horticultural traits in single cross hybrids of cabbage (Brassica oleracea L. var. capitata L.). Indian Journal of Agricultural Sciences 79:703-708
  • Thakur H, Vidyasagar Vidyasagar2016a Estimates of genetic variability, heritability and genetic advances for yield and horticultural traits in cabbage (Brassica oleracea L. var. capitata L.). Journal of Environment and Bio-Sciences 30:155-157
  • Thakur H, Vidyasagar Vidyasagar2016b Combining ability, gene action and heterosis studies involving SI and CMS lines and testers in cabbage. Green Farming 7:580-585
  • Thamburaj S, Singh N2001 Textbook of vegetables, tuber crops and spices. ICAR Publication, New Delhi, 99p
  • Walters TW, Mutschler MA, Earle ED1992 Protoplast fusion-derived Ogura male sterile cauliflower with cold tolerance. Plant Cell Reports 10:624-628

Publication Dates

  • Publication in this collection
    20 Mar 2023
  • Date of issue
    2023

History

  • Received
    21 Oct 2022
  • Accepted
    09 Jan 2023
  • Accepted
    22 Jan 2023
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