陳威臣1 曹進義2 吳姿穎3 夏奇鈮4,*
摘要
陳威臣、曹進義、吳姿穎、夏奇鈮。2021。根莖直徑與兩階段連續培養方式對薑組培苗增
殖與生長之影響。台灣農業研究70(2):117–128。
本研究以「廣東薑」(Zingiber officinale ‘Guang Dong’) 與「竹薑」(Z. officinale ‘Chu’) 組織培養根莖進行試驗，
探討根莖直徑與液、固態兩階段連續培養方式對組培苗增殖與生長之影響。將直徑分別約2、4 及 6 mm 根
莖接種於含有1.0 mg L-1苯甲基腺嘌呤 (benzylaminopurine; BA) 與 0.1 mg L-1萘乙酸 (α-naphthalene acetic acid;
NAA) 之 Murashige & Skoog (MS) 固態培養基培養 8 wk，結果顯示「廣東薑」每一根莖可得 6.2–6.7 芽，但 3
種根莖直徑之間並無顯著差異。「竹薑」4 mm 與 6 mm 之每一根莖可得約 6.8–6.9 芽，顯著高於 2 mm 根莖所
得之芽體；同時考慮瓶苗之株高與鮮重表現，則「廣東薑」與「竹薑」瓶苗增殖皆以利用直徑達4 mm 或以上
之根莖較佳。液、固態兩階段連續培養，係先將直徑約5 mm 之根莖培養於含有 0.1 mg L-1 NAA，並分別添
加0、0.5、1.0、2.0 mg L-1 BA 之 MS 液態培養基中，培養 2、4 或 6 wk 後，將所得之芽苗團塊直接接種於含 有0.5 mg L-1 BA 與 0.1 mg L-1 NAA 之 MS 固態培養基中再培養 8 wk。結果顯示，「廣東薑」以含有 2.0 mg L-1 BA 經 6 wk 液態培養處理之 24.6 芽顯著最多；「竹薑」在 6 wk 液態培養之 0.5–2.0 mg L-1 BA 處理皆具有最
高總芽數。綜合上述結果，建議利用直徑4 mm 以上之根莖作為培植體，可兼顧瓶苗之增殖效率與生長品質；
液、固態兩階段連續培養試驗中，若依平均每週可生產之芽數來看，「廣東薑」以2.0 mg L-1 BA 處理與「竹薑」
以1.0–2.0 mg L-1 BA 處理之根莖，於液態培養 4 wk 後再繼代培養於固態培養基繼續培養 8 wk，所得之總芽
數為最高。
關鍵詞：薑、微體繁殖、組培根莖、液態培養、兩階段連續培養。
收件日期：2020 年 12 月 2 日；接受日期：2021 年 2 月 17 日。
* 通訊作者：[email protected]
1 農委會農業試驗所生物技術組助理研究員。台灣 台中市。
2 農委會農業試驗所生物技術組聘用助理研究員。台灣 台中市。
3 農委會農業試驗所生物技術組計畫助理。台灣 台中市。
4 農委會農業試驗所生物技術組研究員。台灣 台中市。
前言
薑 (Zingiber officinale) 為薑科 (Zingiber-aceae) 多年生宿根草本根莖類植物，原生於東 南亞熱帶地區，因其根莖 (rhizome) 兼具食用 與藥用功能，自古應用於世界各地。薑除了應 用在食物調理與藥膳食療外，傳統醫學常用於 緩解或治療風濕關節炎、扭傷酸痛、腸胃不適 及發燒頭痛等症狀 (Ali et al. 2008; Deng & Pan
2009; Rehman et al. 2011; Mishra et al. 2012;
Singh et al. 2014)。現代藥理學亦證實，薑具 有清熱活血、延緩衰老、護肝保胃及改善血液 循環等功能，同時具有降血糖、降血脂和降膽 固 醇 的 效 果， 以 及 抗 菌、 抗 發 炎、 抗 氧 化 及 抗 癌 等 效 用 (Singh et al. 2014; Zadeh & Kor 2014; Agrahari et al. 2015)。
台灣生薑的主要產區在南投、台東及苗栗 等地，種植之品種多為「廣東薑」 (Z. officinale
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118 台灣農業研究 第70 卷 第 2 期
‘Guang Dong’)， 少 量 為「竹 薑」(Z. officinale
‘Chu’)。生薑依其生育期長短，可分為嫩薑、
粉 薑、 老 薑 (或 稱 薑 母 ) 等 3 種 (Agriculture and Food Agency 2017; Council of Agriculture 2017a, 2017b; Chen et al. 2018)。薑栽培以無 性 繁 殖 根 莖 為 主， 但 種 薑 因 為 常 感 染 薑 軟 腐 病 菌 (Pythium myriotylum) 與 薑 青 枯 病 菌 (Ralstonia solanacearum) 而 品 質 下 降。 上 述 病菌係藉由土壤或種薑而擴大傳播，因此連作 薑田之病害發生率高，嚴重影響生薑的產量與 品質 (Kambaska & Santilata 2009; Kavyashree 2009; Sathyagowri & Seran 2011)。 此 外， 由 於種薑不易長期貯存，栽種適期常面臨種薑缺
Ma & Gang 2006; Deng & Pan 2009; Lincy &
Sasikumar 2010; Liu et al. 2010; Seran 2013;
Li 2016; Kasilingam et al. 2018)。利用液態培 養或是液、固態兩階段連續培養方式，可提高 組培薑苗增殖與生長效率 (Lincy & Sasikumar 2010; Hapsari et al. 2011)。本研究前期已建立
「廣東薑」與「竹薑」微體繁殖流程，並以液態
(Bacto Agar, Becton Dickinson and Company, Sparks, MD, USA) 前，先以 0.1–1.0 N NaOH (orbital shaker, SanKuan Co., Taichung, Tai-wan) 以 80 rpm 速度進行振盪培養。
根莖直徑對薑組培苗增殖與生長之影響 選取根莖直徑分別約為2、4 或 6 mm 之「廣 東薑」與「竹薑」組培苗，切除根系但保留2.5 cm 長之假莖作為培植體，接種於內含 100 mL 培養基之500-mL 蘭花瓶 (orchid flask, Taiwan Glass Ind. Corp., Hsinchu, Taiwan) 中 進 行 試 驗。 培 養 基 為 含 有1.0 mg L-1苯 甲 基 腺 嘌 呤 [benzylamino-purine (BA), Sigma, St Louis, MO, USA] 與 0.1 mg L-1萘乙酸 [α-naphthalene acetic acid (NAA), Sigma, St Louis, MO, USA] 之Murashige
& Skoog (MS) 固態培養基 (Murashige & Skoog 1962)。培養 8 wk 後，調查每一培植體所得之
液、固態連續培養方式與BA 濃度對薑 組培苗增殖與生長之影響
將上述不同液態培養時間與BA 濃度之組 合試驗所得之芽苗團塊作為培植體，直接接種 於內含100 mL 固態培養基之 500-mL 蘭花瓶 中。培養基為含有0.5 mg L-1 BA 與 0.1 mg L-1 NAA 之 MS 固態培養基 (簡稱為生長培養基)，
經8 wk 培養後，調查每一培植體所得之芽數。
本 試 驗 每 處 理 採3 重複，每重複 (瓶 ) 接種 3 個培植體。
試驗設計和統計分析
本 研 究 採 用 完 全 逢 機 設 計 (completely randomized design; CRD) 進行試驗。試驗所 得若為百分率資料，則先經角度轉換後再進行 分 析， 經SAS Enterprise Guide 7.1 套裝統計 分析軟體進行變方分析 (analysis of variance;
ANOVA)。若處理間差異顯著 (P < 0.05)，則 以 最 小 顯 著 差 異 性 測 驗 (least significant dif-ference test; LSD test) 比較各處理平均值間之 差異。
(A)
(C)
(E)
(B)
(D)
(F)
圖1. 「竹薑」根莖於含有不同濃度 BA 之液態培養基中，培養不同時間後之叢生芽苗團塊或組培苗形成情
形。以直徑約5 mm 根莖培養於含有 0.1 mg L-1 萘乙酸 (α-naphthalene acetic acid; NAA) 之 Murashige & Skoog (MS) 液態培養基，並添加 (A) 0、(B) 1 mg L-1 苯甲基腺嘌呤 (benzyl-aminopurine; BA) 培養 2 wk，(C) 0.5、(D) 2 mg L-1 BA 培養 4 wk，以及 (E) 0.5、(F) 1 mg L-1 BA 培養 6 wk。YS：黃色芽、GS：綠色芽、PL：葉片展開 芽。Bars = 1 cm。
Fig. 1. Shoot-bud clusters and plantlets of in vitro Zingiber offi cinale ‘Chu’ derived from media with various ben-zylamino-purine (BA) concentrations and culture durations. In vitro rhizomes (about 5 mm in diameter) were cultured in a MS liquid medium supplemented with 0.1 mg L-1 NAA and in combined with (A) 0, (B) 1.0 mg L-1 BA for 2 wk;
(C) 0.5, (D) 2.0 mg L-1 BA for 4 wk and (E) 0.5, (F) 1.0 mg L-1 BA for 6 wk of culture. YS: yellow shoot-bud; GS:
green shoot-bud; and PL: plantlet with unfolded leave. Bars = 1 cm.
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120 台灣農業研究 第70 卷 第 2 期
結果
根莖直徑對薑組培苗增殖與生長之影響 將直徑約為2、4 或 6 mm 之「廣東薑」與
「竹薑」根莖，接種於含有1.0 mg L-1 BA 與 0.1 mg L-1 NAA 之 MS 固態培養基中，培養 8 wk 後 之 結 果 如 表1 與圖 2。「廣東薑」每一根莖 所 得 芽 數 在6.2–6.7 之間，3 種根莖直徑間並
表1. 「廣東薑」與「竹薑」根莖直徑對組培苗增殖與生長之影響。
Table 1. Effect of rhizome diameter on proliferation rate and growth of in vitro Zingiber offi cinale ‘Guang Dong’
and ‘Chu’ plantletsz.
Cultivar Rhizome diameter
(mm) Total plantlets/explant
(number) Plantlet height
(cm) Fresh weight/plantlet (g)
‘Guang Dong’ 2 6.2 ± 0.3 ay 2.6 ± 0.2 b 3.1 ± 0.2 b
4 6.6 ± 0.1 a 3.0 ± 0.2 ab 4.9 ± 0.3 a
6 6.7 ± 0.0 a 3.5 ± 0.1 a 5.3 ± 0.1 a
‘Chu’ 2 4.9 ± 0.2 b 2.5 ± 0.1 b 2.5 ± 0.2 b
4 6.8 ± 0.3 a 3.2 ± 0.2 a 4.4 ± 0.2 a
6 6.9 ± 0.1 a 3.1 ± 0.3 ab 4.6 ± 0.5 a
zIn vitro rhizome explants (about 2, 4 or 6 mm in diameter) were cultured on Murashige & Skoog (MS) medium containing 1.0 mg L-1 benzyl-aminopurine (BA) and 0.1 mg L-1 α-naphthalene acetic acid (NAA) for 8 wk of culture. Each treatment had 3 repli-cations, and 3 explants were used in each replication. Plantlets higher than 1 cm were recorded.
y Means in the same column in each cultivar followed by different letters are signifi cantly different at the 5% level by least signifi -cant difference (LSD) test.
(A1) (A2) (A3)
(B1) (B2) (B3)
圖2. 利用不同直徑薑根莖培養所得之組培苗增殖與生長情形。利用根莖直徑約為 (A1、B1) 2 mm、(A2、
B2) 4 mm 及 (A3、B3) 6 mm 之 (A1–A3)「廣東薑」與 (B1–B3)「竹薑」根莖培植體，培養於含有 1.0 mg L-1 苯甲基腺嘌呤 (benzyl-aminopurine; BA) 與 0.1 mg L-1萘乙酸 (α-naphthalene acetic acid; NAA) 之 Murashige &
Skoog (MS) 固態培養基中，培養 8 wk 後之組培苗增殖與生長情形。Bar = 2 cm。
Fig. 2. Proliferation and growth of in vitro Zingiber offi cinale plantlets were obtained by using rhizome in different diameters. In vitro plantlets of (A1–A3) Z. offi cinale ‘Guang Dong’ and (B1–B3) ‘Chu’ derived from rhizomes with about (A1, B1) 2, (A2, B2) 4, and (A3, B3) 6 mm in diameter were cultured on a MS medium supplemented 1.0 mg L-1 BA and 0.1 mg L-1 NAA for 8 wk of culture. Bar = 2 cm.
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無顯著差異。苗高以6 mm 根莖所得芽體之 3.5
122 台灣農業研究 第70 卷 第 2 期
表2. 液、固態兩階段連續培養與BA濃度對「廣東薑」組織培養苗增殖與生長之影響。 Table 2. Effects of two-stage continuous culture and BA concentration on proliferation rate and growth of in vitro Zingiber officinale ‘Guang Dong’ plantlets.
Liquid culture period (wk)
BA (mg L-1)
Liquid culturezSolid culture (8 wk)y Total plantlets/ explant
Plantlets with yellowish buds
(%)
Plantlets with green shoots
(%)
Plantlets with unfolded leaves
(%)Total plantlets/ expla
Plantlets with green shoots
(%)
Plantlets with unfolded leaves
(%) 20.0 3.8 ± 0.2 gx 100.0 a0.0 c0.0 d8.9 ± 1.8 e 21.8 ± 6.9 a78.2 ± 6.9 c 0.54.2 ± 0.3 fg100.0 a0.0 c0.0 d 9.8 ± 1.3 de 19.9 ± 4.6 ab80.1 ± 4.6 c 1.04.2 ± 0.4 fg100.0 a0.0 c0.0 d 9.8 ± 2.0 de 13.5 ± 2.6 ab 86.5 ± 2.6 bc 2.04.6 ± 0.3 ef100.0 a0.0 c0.0 d 9.7 ± 0.6 de 14.6 ± 5.1 ab 85.4 ± 5.1 bc 40.0 5.1 ± 0.2 cde47.2 ± 16.0 b52.8 ± 16.0 a0.0 d13.0 ± 1.5 c9.1 ± 4.7 b90.9 ± 4.7 b 0.5 5.3 ± 0.1 bcd65.5 ± 18.6 b34.5 ± 18.6 ab0.0 d20.5 ± 1.3 b 10.1 ± 5.1 ab 89.9 ± 5.1 bc 1.0 5.2 ± 0.1 bcde100.0 a0.0 c0.0 d20.1 ± 1.6 b 11.7 ± 2.1 ab 88.3 ± 2.1 bc 2.05.0 ± 0.2 de100.0 a0.0 c0.0 d20.0 ± 2.8 b 10.0 ± 2.4 ab 90.0 ± 2.4 bc 60.0 5.7 ± 0.1 abc11.8 ± 11.8 c37.9 ± 4.7 ab50.3 ± 7.7 a 12.4 ± 1.8 cd0.0 c100.0 a 0.56.1 ± 0.4 a50.2 ± 10.7 b27.4 ± 10.5 b22.4 ± 0.4 b19.6 ± 2.4 b0.0 c100.0 a 1.05.8 ± 0.1 ab50.3 ± 9.2 b28.9 ± 10.2 ab20.8 ± 11.3 bc21.1 ± 1.3 b 12.8 ± 1.6 ab 87.2 ± 1.6 bc 2.05.8 ± 0.2 ab54.6 ± 19.0 b38.0 ± 14.4 ab7.4 ± 4.9 c24.6 ± 1.7 a 13.1 ± 4.6 ab 86.9 ± 4.6 bc SourceF-testw Wk********************* BAns******** nsns Wk × BAns********** z In vitro rhizome explants (about 5 mm in diameter) were cultured on Murashige & Skoog (MS) liquid medium containing 0.0–2.0 mg L-1 benzyl-aminopurine (BA) and 0.1 mg L-1 α-naphthalene acetic acid (NAA) for 2, 4 and 6 wk of culture. Each treatment had 3 replications, and 3 explants were used in each replication. y Uncut shoot-bud derived from various treatments in liquid medium were subcultured in the MS solid medium supplemented with 0.5 mg L-1 BA and 0.1 mg L-1 NAA for extending 8 wk of culture. x Means in each same column followed by different letters are significantly different at the 5% level by least significant difference (LSD) test. Percentage data were arcsine-square-root transformed prior to analysis. w F-test of analysis of variance (ANOVA). ns: non-significant; *, ** and ***, are significant at 5%, 1% and 0.1% level, respectively.
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表3. 液、固態兩階段連續培養與BA濃度對「竹薑」組織培養苗增殖與生長之影響。 Table 3. Effects of two-stage continuous culture and BA concentration on proliferation rate and growth of in vitro Zingiber officinale ‘Chu’ plantlets.
Liquid culture period (wk)
BA (mg L-1)
Liquid culturezSolid culture (8 wk)y Total plantlets/ explant
Plantlets with yellowish buds
(%)
Plantlets with green shoots
(%)
Plantlets with unfolded leaves
(%)Total plantlets/ explant
Plantlets with green shoots
(%)
Plantlets with unfolded leaves
(%) 20.04.0 ± 0.2 ex 100.0 a0.0 c0.0 c4.6 ± 0.1 d0.0 e100.0 a 0.54.3 ± 0.0 de100.0 a0.0 c0.0 c5.4 ± 0.2 cd9.4 ± 2.3 ab90.6 ± 2.3 de 1.04.3 ± 0.3 de100.0 a0.0 c0.0 c6.2 ± 0.6 cd7.7 ± 2.4 ab92.3 ± 2.4 de 2.04.2 ± 0.5 e100.0 a0.0 c0.0 c6.1 ± 1.1 cd12.9 ± 1.4 a87.1 ± 1.4 e 40.05.3 ± 0.2 cd100.0 a0.0 c0.0 c5.9 ± 0.5 cd3.2 ± 3.2 cde96.8 ± 3.2 abc 0.55.8 ± 0.3 bc92.6 ± 7.4 ab7.4 ± 7.4 bc0.0 ± 0.1 c8.0 ± 1.7 bc4.7 ± 2.5 bcd95.3 ± 2.5 bcd 1.05.9 ± 0.1 bc88.5 ± 8.4 b11.5 ± 8.4 b0.0 ± 0.1 c10.3 ± 1.0 ab3.2 ± 3.2 cde96.8 ± 3.2 abc 2.05.9 ± 0.1 bc87.3 ± 2.3 b12.7 ± 2.3 ab0.0 c11.1 ± 1.2 a5.4 ± 4.1 abc94.6 ± 4.1 cde 60.05.4 ± 0.1 bc5.9 ± 0.4 d23.7 ± 5.8 a70.4 ± 5.8 a7.2 ± 0.3 cd0.0 e100.0 a 0.55.9 ± 0.1 bc32.2 ± 4.9 c26.9 ± 3.3 a40.9 ± 4.2 b10.4 ± 0.6 ab1.5 ± 1.1 cde98.5 ± 1.1 abc 1.06.4 ± 0.2 ab30.4 ± 8.6 c25.8 ± 6.4 a43.8 ± 5.6 b10.3 ± 0.8 ab0.0 e100.0 a 2.07.0 ± 0.6 a32.0 ± 7.3 c28.7 ± 2.9 a39.4 ± 6.8 b10.7 ± 0.2 ab0.9 ± 0.9 de99.1 ± 0.9 ab SourceF-testw Wk********************* BAnsnsns********* Wk × BAns*ns***nsnsns z In vitro rhizome explants (about 5 mm in diameter) were cultured on Murashige & Skoog (MS) liquid medium containing 0–2.0 mg L-1 benzyl-aminopurine (BA) and 0.1 mg L-1 α-naphthalene acetic acid (NAA) for 2, 4 and 6 wk of culture. Each treatment had 3 replications, and 3 explants were used in each replication. y Uncut shoot-bud derived from various treatments in liquid medium were subcultured in the MS solid medium supplemented with 0.5 mg L-1 BA and 0.1 mg L-1 NAA for extending 8 wk of culture. x Means in each same column followed by different letters are significantly different at the 5% level by least significant difference (LSD) test. Percentage data were arcsine-square-root transformed prior to analysis. w F-test of analysis of variance (ANOVA). ns: non-significant; *, ** and ***, are significant at 5%, 1% and 0.1% level, respectively.
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萱草 (Hemerocallis spp.) (Chen et al. 2005)、
海 棗 (Phoenix dactylifera) (Mazri 2013) 及朝 鮮薊 (Cynara cardunculus) (El Boullani et al.
2017) 研究則指出，組織培養各階段均有其適 用培植體大小，過大或太小培植體均不利於組 培 苗 生 長。 然 而，Chen et al. (2019) 於 孤 挺 花 (Hippeastrum hybridum) 研究結果顯示，培 植體來源之鱗莖大小對組培苗增殖並無顯著影 Sathyagowri & Seran (2011) 在薑的研究相符，
即較長的新生芽體培植體 (2.0 cm)，應當具有
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