生物技术通报 ›› 2026, Vol. 42 ›› Issue (1): 105-113.doi: 10.13560/j.cnki.biotech.bull.1985.2025-0833
樊荣辉(
), 罗远华, 陈艺荃, 方能炎, 陈燕, 钟淮钦, 叶秀仙(
)
收稿日期:2025-08-02
出版日期:2026-01-26
发布日期:2026-02-04
通讯作者:
叶秀仙,女,研究员,研究方向 :花卉组织培养;E-mail: yxx7861@163.com作者简介:樊荣辉,女,副研究员,研究方向 :花卉分子育种;E-mail: rhfan1012@163.com
基金资助:
FAN Rong-hui(
), LUO Yuan-hua, CHEN Yi-quan, FANG Neng-yan, CHEN Yan, ZHONG Huai-qin, YE Xiu-xian(
)
Received:2025-08-02
Published:2026-01-26
Online:2026-02-04
摘要:
目的 从南茜组培苗中选育出新株系‘金辉’,株型高大,花枝挺立,但无香气。通过解析无香‘金辉’和浓香‘香水文心’花香形成的分子机制,探索‘金辉’无香气制约因素,以期通过分子技术使‘金辉’释放香气。 方法 通过顶空-固相微萃取气相色谱法(HS-SPME)/GC-MS测定文心兰2个品种香气成分及含量。以‘金辉’和‘香水文心’不同发育时期花为材料,通过转录组测序和荧光定量PCR验证,筛选花香形成关键基因。 结果 气相色谱-质谱分析表明,‘香水文心’盛花期花香挥发物总含量显著高于‘金辉’,是‘金辉’的6.1倍,‘香水文心’花香挥发物以单萜为主,‘金辉’花香挥发物以倍半萜为主。转录组测序结果表明,共获得55 277个unigenes,大约81.64%的unigenes被公共数据库注释。单萜形成的上游途径,MEP途径中,多数基因,如CMK、HDR、HDS、GPPS等在‘香水文心’中表达高于‘金辉’,使生成单萜的前体物质GPP在‘香水文心’中积累。倍半萜形成的上游途径,MVA途径中,约一半基因在‘香水文心’中略高表达,一半基因在‘金辉’中略高表达。转录组数据中共筛选出3个显著差异表达的萜类合成酶基因(TPS)。其中,OhTPS1和OhTPS2属于TPS-b家族,主要由单萜合成酶(mono-TPSs)基因组成。OhTPS1 和OhTPS2在‘香水文兰’中表达显著高于‘金辉’中,且在‘香水文心’中随着花的发育逐渐升高,表明这2个基因在‘香水文心’单萜生物合成中起关键作用。OhTPS3属于TPS-a家族,主要由倍半萜合成酶(sesqui-TPSs)基因组成。OhTPS3 在‘金辉’中表达显著高于‘香水文心’中,且在‘金辉’中随着花的发育逐渐升高,表明这个基因在‘金辉’倍半萜生物合成中起关键作用。 结论 ‘金辉’花香挥发物总量低的主要原因是MEP途径多数关键基因低表达,一定程度限制代谢流向单萜前体物质GPP积累,同时单萜合成酶基因的低表达,也一定程度阻碍单帖挥发物的生成。
樊荣辉, 罗远华, 陈艺荃, 方能炎, 陈燕, 钟淮钦, 叶秀仙. 文心兰‘金辉’和‘香水文心’花香形成比较分析[J]. 生物技术通报, 2026, 42(1): 105-113.
FAN Rong-hui, LUO Yuan-hua, CHEN Yi-quan, FANG Neng-yan, CHEN Yan, ZHONG Huai-qin, YE Xiu-xian. Comparative Analysis of Floral Scent Formation between ‘Jinhui’ and Onc. ‘Sharry Baby’ of Oncidium hybridum[J]. Biotechnology Bulletin, 2026, 42(1): 105-113.
基因 Gene | 正向引物 Forward primer sequence (5′-3′) | 反向引物 Reverse primer sequence (5′-3′) |
|---|---|---|
| OhTPS1 | ATGATGACTACATACAATCGCTAC | TCAAGTCCAAGTTGGCATAG |
| OhTPS2 | ACCTCGTCTACATCATCAGC | CCTTCAATTTATTCCTTTCG |
| OhTPS3 | TTATTGATGCGCTACAACG | CAATCCACGTACATTTCCA |
| Actin | AATGTGCCTGCTATGTATGTTGCT | GGGCATATCCTTCGTAGATTG |
表1 RT-qPCR所用引物序列
Table 1 Primer sequences for RT-qPCR
基因 Gene | 正向引物 Forward primer sequence (5′-3′) | 反向引物 Reverse primer sequence (5′-3′) |
|---|---|---|
| OhTPS1 | ATGATGACTACATACAATCGCTAC | TCAAGTCCAAGTTGGCATAG |
| OhTPS2 | ACCTCGTCTACATCATCAGC | CCTTCAATTTATTCCTTTCG |
| OhTPS3 | TTATTGATGCGCTACAACG | CAATCCACGTACATTTCCA |
| Actin | AATGTGCCTGCTATGTATGTTGCT | GGGCATATCCTTCGTAGATTG |
| 序号No. | 保留时间Retention time (min) | 挥发化合物 Volatile component | 分子式 Molecular formula | 含量 Volatile abundance (μg/g) | |||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| 金辉 Jinhui | 香水文心Onc. Sharry baby | ||||||||||
| 花蕾前期Early bud period | 花蕾中期Mid bud period | 盛花期Blooming period | 花蕾前期Early bud period | 花蕾中期Mid bud period | 盛花期Blooming period | ||||||
| 1 | 1.931 | 丙酮 2-Propanone | C3H6O | 0.11±0.02 | 0.17±0.02 | 0.01±0.01 | - | - | - | ||
| 2 | 2.833 | 3-戊酮 3-Pentanone | C5H10O | 0.27±0.05 | 0.29±0.03 | 0.11±0.03 | - | - | - | ||
| 3 | 2.921 | 叶醇 Leaf alconol | C6H12O | - | - | - | 0.32±0.07 | 0.62±0.07 | 0.65±0.07 | ||
| 4 | 3.933 | 正己醛 Hexanal | C6H12O | 0.35±0.04 | 0.23±0.03 | 0.05±0.02 | 0.64± 0.33 | 0.74±0.38 | - | ||
| 5 | 4.427 | 正己醇 Hexyl alconol | C6H14O | - | - | - | 0.33±0.05 | 0.58±0.05 | 1.08±0.07 | ||
| 6 | 5.233 | 丙烯醛 2-Propenal | C7H6O2 | - | - | - | 0.56±0.43 | 0.43±0.23 | - | ||
| 7 | 5.876 | α-派烯 α-Pinene | C10H16 | - | - | - | - | 1.56±0.54 | 6.47±0.45 | ||
| 8 | 6.746 | 1-戊烯-3-醇 Ethyl vinyl carbinol | C7H9ClO | 0.15±0.03 | 0.30±0.03 | 0.05±0.02 | - | - | - | ||
| 9 | 7.500 | 香桧烯(+)-Sabinene | C10H16 | - | - | - | - | 0.12±0.01 | 0.55±0.09 | ||
| 10 | 7.672 | β-派烯 β-Pinene | C10H16 | - | - | - | - | 0.20±0.02 | 0.44±0.12 | ||
| 11 | 8.245 | β-月桂烯 β-Myrcene | C10H16 | - | - | - | 1.83±0.13 | 6.59±0.30 | 6.50±0.63 | ||
| 12 | 9.834 | D-柠檬烯 D-Limonene | C10H16 | - | - | - | - | 0.62±0.12 | 2.74±0.22 | ||
| 13 | 9.928 | 1,8-桉叶素 1,8-Cineole | C10H16 | - | - | - | - | 6.27±0.63 | 15.34±0.79 | ||
| 14 | 10.080 | 苯甲醇 Benzyl alcohol | C7H8O | - | - | - | - | 0.62±0.16 | 2.37±0.83 | ||
| 15 | 10.616 | β-罗勒烯 β-Ocimene | C10H16 | - | 0.44±0.02 | 1.83±0.13 | - | 10.39±0.51 | 31.84±1.47 | ||
| 16 | 12.784 | 芳樟醇 Linalool | C10H16 | - | - | - | - | 2.34±0.23 | 10.63±0.56 | ||
| 17 | 14.912 | 异辛醇 Isooctanol | C8H18O | - | - | - | 0.51±0.23 | 0.52±0.15 | - | ||
| 18 | 15.567 | 顺-3-壬烯醇 cis-3-Nonenol | C9H18O | - | - | - | - | - | 0.30±0.05 | ||
| 19 | 15.953 | γ-木糖烯 γ-Muurolene | C15H24 | - | 0.20±0.01 | 0.10±0.04 | - | - | - | ||
| 20 | 16.383 | α-萜品烯 α-Terpineol | C10H18O | - | - | - | 1.92±0.25 | 7.08±2.88 | |||
| 21 | 16.425 | α-古巴烯 α-Copaene | C15H24 | 0.12±0.02 | 1.54±0.23 | 8.49±0.41 | - | - | - | ||
| 22 | 17.155 | β-波旁烯 β-Bourbonene | C15H24 | - | 0.07±0.02 | 0.25±0.04 | - | 0.41±0.10 | 1.30±0.17 | ||
| 23 | 20.807 | γ-榄香烯 γ-Elemene | C15H24 | - | 0.04±0.01 | 0.14±0.03 | - | - | - | ||
| 24 | 21.029 | 1-十六炔 1-Hexadecyne | C15H24 | - | - | - | - | - | 0.50±0.10 | ||
| 25 | 21.447 | α-古芸烯 α-Gurjunene | C15H24 | - | 0.02±0.01 | 0.16±0.02 | - | - | - | ||
| 26 | 23.121 | β-瑟林烯 β-Selinene | C15H24 | - | - | 0.12±0.03 | - | - | - | ||
| 27 | 23.220 | β-橄榄烯 γ-Maaliene | C15H24 | - | 0.10±0.02 | 0.30±0.05 | - | - | - | ||
| 28 | 24.179 | β-石竹烯 β-Caryophyllene | C15H24 | - | 0.72±0.05 | 2.94±0.15 | - | 0.10±0.06 | 2.03±0.08 | ||
| 29 | 25.955 | γ-杜松烯 γ-Cadinene | C15H24 | - | - | - | - | 0.12±0.03 | 0.66±0.06 | ||
| 30 | 26.140 | 大根香叶烯 Germacrene D | C15H24 | - | 0.03±0.01 | 0.11±0.07 | - | 0.55± 0.04 | 4.58±1.61 | ||
| 31 | 26.611 | 惕各酸苄酯 Benzyl tiglate | C12H14O2 | - | - | - | - | 0.31±0.06 | 0.89±0.30 | ||
| 32 | 26.846 | α-法呢烯 α-Farnesene | C15H24 | - | 0.09±0.01 | 0.05±0.05 | - | 2.06±0.51 | 10.30±1.09 | ||
| 33 | 28.573 | 反式-橙花叔醇 Nerolidol | C15H24 | - | - | - | - | 0.37± 0.06 | 1.60±0.20 | ||
| 34 | 28.921 | 鲨烯 Squalene | C30H50 | - | - | - | - | 1.28±0.36 | 4.98±0.42 | ||
表2 不同文心兰品种主要花香成分
Table 2 Main floral components of different varieties in O. hybridum
| 序号No. | 保留时间Retention time (min) | 挥发化合物 Volatile component | 分子式 Molecular formula | 含量 Volatile abundance (μg/g) | |||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| 金辉 Jinhui | 香水文心Onc. Sharry baby | ||||||||||
| 花蕾前期Early bud period | 花蕾中期Mid bud period | 盛花期Blooming period | 花蕾前期Early bud period | 花蕾中期Mid bud period | 盛花期Blooming period | ||||||
| 1 | 1.931 | 丙酮 2-Propanone | C3H6O | 0.11±0.02 | 0.17±0.02 | 0.01±0.01 | - | - | - | ||
| 2 | 2.833 | 3-戊酮 3-Pentanone | C5H10O | 0.27±0.05 | 0.29±0.03 | 0.11±0.03 | - | - | - | ||
| 3 | 2.921 | 叶醇 Leaf alconol | C6H12O | - | - | - | 0.32±0.07 | 0.62±0.07 | 0.65±0.07 | ||
| 4 | 3.933 | 正己醛 Hexanal | C6H12O | 0.35±0.04 | 0.23±0.03 | 0.05±0.02 | 0.64± 0.33 | 0.74±0.38 | - | ||
| 5 | 4.427 | 正己醇 Hexyl alconol | C6H14O | - | - | - | 0.33±0.05 | 0.58±0.05 | 1.08±0.07 | ||
| 6 | 5.233 | 丙烯醛 2-Propenal | C7H6O2 | - | - | - | 0.56±0.43 | 0.43±0.23 | - | ||
| 7 | 5.876 | α-派烯 α-Pinene | C10H16 | - | - | - | - | 1.56±0.54 | 6.47±0.45 | ||
| 8 | 6.746 | 1-戊烯-3-醇 Ethyl vinyl carbinol | C7H9ClO | 0.15±0.03 | 0.30±0.03 | 0.05±0.02 | - | - | - | ||
| 9 | 7.500 | 香桧烯(+)-Sabinene | C10H16 | - | - | - | - | 0.12±0.01 | 0.55±0.09 | ||
| 10 | 7.672 | β-派烯 β-Pinene | C10H16 | - | - | - | - | 0.20±0.02 | 0.44±0.12 | ||
| 11 | 8.245 | β-月桂烯 β-Myrcene | C10H16 | - | - | - | 1.83±0.13 | 6.59±0.30 | 6.50±0.63 | ||
| 12 | 9.834 | D-柠檬烯 D-Limonene | C10H16 | - | - | - | - | 0.62±0.12 | 2.74±0.22 | ||
| 13 | 9.928 | 1,8-桉叶素 1,8-Cineole | C10H16 | - | - | - | - | 6.27±0.63 | 15.34±0.79 | ||
| 14 | 10.080 | 苯甲醇 Benzyl alcohol | C7H8O | - | - | - | - | 0.62±0.16 | 2.37±0.83 | ||
| 15 | 10.616 | β-罗勒烯 β-Ocimene | C10H16 | - | 0.44±0.02 | 1.83±0.13 | - | 10.39±0.51 | 31.84±1.47 | ||
| 16 | 12.784 | 芳樟醇 Linalool | C10H16 | - | - | - | - | 2.34±0.23 | 10.63±0.56 | ||
| 17 | 14.912 | 异辛醇 Isooctanol | C8H18O | - | - | - | 0.51±0.23 | 0.52±0.15 | - | ||
| 18 | 15.567 | 顺-3-壬烯醇 cis-3-Nonenol | C9H18O | - | - | - | - | - | 0.30±0.05 | ||
| 19 | 15.953 | γ-木糖烯 γ-Muurolene | C15H24 | - | 0.20±0.01 | 0.10±0.04 | - | - | - | ||
| 20 | 16.383 | α-萜品烯 α-Terpineol | C10H18O | - | - | - | 1.92±0.25 | 7.08±2.88 | |||
| 21 | 16.425 | α-古巴烯 α-Copaene | C15H24 | 0.12±0.02 | 1.54±0.23 | 8.49±0.41 | - | - | - | ||
| 22 | 17.155 | β-波旁烯 β-Bourbonene | C15H24 | - | 0.07±0.02 | 0.25±0.04 | - | 0.41±0.10 | 1.30±0.17 | ||
| 23 | 20.807 | γ-榄香烯 γ-Elemene | C15H24 | - | 0.04±0.01 | 0.14±0.03 | - | - | - | ||
| 24 | 21.029 | 1-十六炔 1-Hexadecyne | C15H24 | - | - | - | - | - | 0.50±0.10 | ||
| 25 | 21.447 | α-古芸烯 α-Gurjunene | C15H24 | - | 0.02±0.01 | 0.16±0.02 | - | - | - | ||
| 26 | 23.121 | β-瑟林烯 β-Selinene | C15H24 | - | - | 0.12±0.03 | - | - | - | ||
| 27 | 23.220 | β-橄榄烯 γ-Maaliene | C15H24 | - | 0.10±0.02 | 0.30±0.05 | - | - | - | ||
| 28 | 24.179 | β-石竹烯 β-Caryophyllene | C15H24 | - | 0.72±0.05 | 2.94±0.15 | - | 0.10±0.06 | 2.03±0.08 | ||
| 29 | 25.955 | γ-杜松烯 γ-Cadinene | C15H24 | - | - | - | - | 0.12±0.03 | 0.66±0.06 | ||
| 30 | 26.140 | 大根香叶烯 Germacrene D | C15H24 | - | 0.03±0.01 | 0.11±0.07 | - | 0.55± 0.04 | 4.58±1.61 | ||
| 31 | 26.611 | 惕各酸苄酯 Benzyl tiglate | C12H14O2 | - | - | - | - | 0.31±0.06 | 0.89±0.30 | ||
| 32 | 26.846 | α-法呢烯 α-Farnesene | C15H24 | - | 0.09±0.01 | 0.05±0.05 | - | 2.06±0.51 | 10.30±1.09 | ||
| 33 | 28.573 | 反式-橙花叔醇 Nerolidol | C15H24 | - | - | - | - | 0.37± 0.06 | 1.60±0.20 | ||
| 34 | 28.921 | 鲨烯 Squalene | C30H50 | - | - | - | - | 1.28±0.36 | 4.98±0.42 | ||
图4 文心兰TPS基因进化树分析AaADS, 黄蒿紫穗槐-4,11-二烯合酶(AFA34434); AaCAS, 黄蒿β-石竹烯合酶(AAL79181); AaFS, 黄蒿β-法呢烯合酶(Q9FXY7); LsGES1, 莴苣大根香叶烯合酶(AAM11626); MpFS, 胡椒薄荷β-法呢烯合酶(AAB95209); AaLIS, 黄蒿芳樟醇合酶(AAF13356); AtTPS10, 拟南芥月桂烯/罗勒烯合酶(AAG09310); ClLS1, 柠檬柠檬烯合酶1(AAM53944)
Fig. 4 Phylogenetic analysis of the TPS in Oncidium hybridumAaADS, Artemisia annua amorpha-4,11-dienesynthase (AFA34434); AaCAS, Artemisia annua β-caryophyllene synthase (AAL79181); AaFS, Artemisia annua β-farnesene synthase (Q9FXY7); LsGES1, Lactuca sativus germacrene A synthase (AAM11626); MpFS, Mentha piperita β-farnesene synthase (AAB95209); AaLIS, Artemisia annua linalool synthase (AAF13356); AtTPS10, Arabidopsis thaliana myrcene/ocimene synthase (AAG09310); ClLS1, Citrus limon limonene synthase 1 (AAM53944)
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