Plant Regeneration via in Vitro Culture of Ovule Obtain by Intergeneric Crossing Between Citrus junos Sieb. et Tanaka and Poncirus trifoliata Raf.

유자와 탱자의 속간교잡후 배주배양에 의한 식물체 유기

  • Published : 1995.11.01

Abstract

As a basic research for breeding new varieties, reciprocal -intergeneric crosses between Citrus junos and P.trifoliata were made. F$_1$ hybrid production using in vitro ovule culture, gametogenesis, and fertilization phenomena were investigated. Frequency of fruit set resulting from crossing of Citrus junos and Poncirus Trifoliata was 16.6% while that of Poncirus Trifoliata and Citrus junos was 11.7%. Callus formation occurred well when ovules at the 6th week after pollination were cultured on MT (Murashige and Tucker) medium supplemented with zeatin 0.5 mg/L and NAA 1.0 or 3.0 mg/L. Immature ovules developed into mature embryos of the MT medium supplemented with 2,4-D 0.1 or 3.0 mg/L. Immature ovules developed into mature embryos of the MT medium supplemented with 2,4 D 0.1 or 0.5 mg/L. The invitro germination rates of 20-week-old ovules set C. junos $\times$ P. Trifoliata and P. Trifoliata $\times$ C. junos were 54.5% and 48.6%, respectively. The emergence ratios of trifoliate hybrids obtained by C. junos $\times$ P. Trifoliata and P. Trifoliata $\times$ C. junos were 56.7% and 100%, respectively. The chromosome number of C. junos and P. Trifoliata was n = 9 or 2n = 18, and the sizes of their pollen grain were 33.75 $\mu$ and 25.0 $\mu$. The length and width of embryo sac in C. junos and P. Trifoliata were 69.38~79.23 $\mu$ and 27.50~38.56 $\mu$, and those of egg cells were 17.50~41.50 $\mu$ and 6.25~8.12$\mu$. Fertilization of C. junos and P. trifoliata terminated 72 h after pollination.

속간교잡에 의한 신품종 육성을 위한 기초 연구로서 유자와 탱자를 정역교잡한 후 배주배양하였고, 자웅배우체 형성 및 수정현상을 조사하였다. 속간교잡의 결과율은 유자$\times$탱자가 16.6%, 탱자$\times$유자는 11.7%이었다. 수분후 6주된 배주를 MT 기본배지에 Zeatin 0.5 mg/L에 NAA 1 또는 3.0 mg/L에 배양했을 때 callus형성율이 높았지만, 14~15주된 배주를 2,4-D 0.1 mg/L 또는 0.5 mg/L이 첨가된 배지에 배양했을 때는 식물체 유기가 잘 되었다. 20주된 배주배양시 발아율은 유자 $\times$ 탱자의 경우 54.5%, 행자 $\times$ 유자는 48.6%이었다. F$_1$식물의 삼엽형 출현율은 유자$\times$탱자는 56.7%, 탱자$\times$유자는 100%이었다. 유자와 행자의 염색체수는 모두 n=9 (2n=18)이었고 화분립의 크기는 각각 33.75 $\mu$, 25.0 $\mu$이었다. 유자와 탱자 배낭의 길이와 폭은 69.38~79.23 $\mu$, 27.50~38.56 $\mu$, 난세포의 길이와 폭은 17.50~41.50 $\mu$, 6.25~8.12 $\mu$이었다. 수분 후 72시간이면 수정이 끝났다.

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