Effects of concentration of inorganic nitrogen, KNos and/or NH4Noj. on growth of embryos in uitro in Petunia hybrida Yoshiji Nrrrr,l Faculty of Agricu

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1 Effects of concentration of inorganic nitrogen, KNos and/or NH4Noj. on growth of embryos in uitro in Petunia hybrida Yoshiji Nrrrr,l Faculty of Agriculture, Nagoga [Jniaersity, Chihusa, Nagoya Summary In oitro culture of just fertilized ovules with placenta of. P. hybri.da. W 166 H X K 146 BH (each of clone is of self-incompatible), was conducted on some inorganic nutrient medium. Normal and abnormal seedlings developed from ovules with placenta at the stage of just fertilized ovules -on MURASHIGE and SKooG's inorganic medium containing large quantities of KNO. and NHaNO!, but no seedlings on other inorganic media. To obtain many normal seedlings, effects of concentration of inorganic nitrogen KNO, and/or NHa- NO3 on the growth of ovules were examined on NttscH's nutrient rhedium. The medium of NITSCH's nutrient plus KNOg 500 ppm and NHaNO3 80 ppm was better for growth of embryo than other media containing high nitrogen salts added to NITSCH's nutri_ ent medium. In conclusion, there were combined effecti' of NO,-N and NHa-N on the growth arrd development i,n aitro of P. hybrida embryo. Nitrogen salt solution for better growth was in a concentration of 12 nm and in ratio of NO'-N to NH.-N in molar unit oi 11:1.

2 Fig. 3. Normal (A) and abnormal (B, C) seedlings developed frorn seeds in P. hybrida on MURASHIGE & Srooc's inorganic nutrient medium. (lnitial samples of ovules with placenta were taken on the 3 rd day after crosspollination of W166HxK146BH in uiuo. Then, seeds obtained from the ovules after 30 days incubation on the MURASIIIGE & Srooc's inorganic nutrient medium. Thereafter, the seeds were sown on Muneslttco & Srooc's medium)

3 .m]:dia-for, ultures of Each media contain ovules with placenta 5/o sucrose and.0.7fr Constituents -Glycine myo-i4ositol Calcium panthothenate Pylidoxiqe. hydrochloride

4 αの胚珠培養に16ける窒素の濃度 と形態 の影響 新美 :& π αり ι 59 咤 ノ 裁 謬 ん ザt プ ` Fig 5 Fig 4 fπ υθdevelopnlent of enlbryo in P/1ノ ll ι j α. fπ γο development of einbryo in Pん υづ b/ αα The ovulcs ー ノ ith placenta on the 3 rd day A 4-ccned proembryo of 10 days, B. Globular vcrc culturcd on after cross p01lination π υづυθ stage proembryo and endosperin of 15 days, and NITSCH's(Ml)and MURASIHGE&SK00G's(M2) C. 20 days after cross nutrient media pollination, K146 BH(e,embryo;en, endosperm). W16611 Longitudinal scctions of 101 days after inoculation sho proembryo on ル ring, 2ヽ 2-cclled 11, B Globular proenlbl yo and ccllular endosperin on N12, and C Abnorinal 胚 乳 の 発 達 は 観 察 され な か つ た が, MURASHIGE& proembryo on A/12(C,enlbryo) S K 0 0 G 培 地 の ものには 細胞 状 の胚 乳 が 発達 してい た π υ υο と比 t 床 2 0 日 後 の両培 地 におけ る胚 発達 は グ 較 して 全1 2 的に遅 れ が 見 られ た N I T s c I I 培地 の胚 珠 に ) は まだ十 分に 分化 していないか 珠か 置床 3 0 日 後 の1 玉 υο とはゃゃ形 態 的 に ' ( な つ た g l o b u l a r s t a g c まj κυ あるいは奇形 と思われ る胚が多数 観察 された ( 第7 メ ) の の胚 が 見 われた が, 胚 乳 は ほ とん ど発達 してい なか つ た ( 第6 図 A ). M U R A S I I I G E & S K 0 0 G 培 地 で培 養 され た B ) さらに M U R A S I H G E & S K 0 0 G 培 NITscIIと MURASIIIGE&SK00G培 地 で培 i [ された 地で そ れ ぞ れ の 胎 座 に つ い たす べ て の 3 0 日 間培 養 した 体 不珠 を たを 行な つ 寒 天培 地 で 発芽 試し 胚 珠 の胚 の形態 は 置床 1 0 日 後 の もの とあ ま り違 わ な い 胎座 か ら切 り聞: し, 1 % の I が, 胚 乳 の発 達 が急 速 に 進 んでい る と思 わ れ た ( 第 6 Ⅸ た 2 0 日 後 には N I T s C I I 培地 で 培 養 され た胚 珠 か らは

5 Fig. 6. In uitro development of embryo in P.hybrida. The ovules with placenta on the 3 rd day after cross-pollination in uiuo were cultured on M, or M2 media. Longitudinal sections of 20 days after inoculation showing, A. Globular stage proembryo on M1 and B. Globular stage proen-rbryo and endosperm on M2 (e, embryo;en,endosperm).

6 Table 3. Ovule culture in P. hfirida, W166HXK146 BH. Effect of the inorganic nitrogen supplemented by KNOg and NHaNO3, separately and in combination, as a function of concentration in NITSCH's nutrient medium (basal medium) on the production of mature seeds and seedlings.

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