Grain Yield, Quality and 2-Acetyl-1-Pyrroline of Fragrant Rice in Response to Different Planting Seasons in South China

Climate conditions is an important factor affected the fragrant rice growth and development.In order to study the effects of different planting seasons on fragrant rice performance in South China,present study was conducted with three planting seasons(early season(April to July),middle season(June to September)and late season(August to November))and three fragrant rice cultivars,‘Basmati-385’,‘Meixiangzhan-2’and‘Xiangyaxiangzhan’.The results showed that the highest grain yield and grain 2-acetyl-1-pyrroline(2-AP,key component of fragrant rice aroma)content were both recorded in late season treatment while the fragrant rice in middle season treatment produced the lowest grain yield,grain filling percentage,1000-grain weight and gain 2-AP content.The highest contents of precursors(proline,pyrroline-5-carboxylic acid and 1-pyrro-line)which related to 2-AP biosynthesis were recorded in late season treatment compared with early season treatment and middle season treatment.The highest activities of enzymes(proline dehydrogenase,pyrroline-5-carboxylic acid synthetase and ornithine transaminase)which involved in 2-AP biosynthesis were also observed in late season treatment.Moreover,the fragrant rice cultivars in late season possessed the lowest chalk rice rate,chalkiness as well as the highest brown rice rate,head rice and protein content.Thus,the optimal season for fragrant rice production in South China is the late season.

Epoxiconazole Improved Photosynthesis,Yield Formation,Grain Quality and 2-Acetyl-1-Pyrroline Biosynthesis of Fragrant Rice

Epoxiconazole is a triazole compound.However,the effects of epoxiconazole on crop productivity and quality were rarely reported.In this study,we investigated the effects of epoxiconazole application on yield formation,grain quality attributes,and 2-acetyl-1-pyrroline(2-AP)content in fragrant rice.A three-year field experiment was carried out with a fragrant rice variety,Meixiangzhan 2.At the heading stage,0,0.02,0.04,0.08,0.16 and 0.32 g/L epoxiconazole solutions were foliar applied to fragrant rice plants,respectively.The results showed that epoxiconazole application significantly increased grain yield,seed-setting rate and 1000-grain weight.Chlorophyll content and net photosynthetic rate of fragrant rice during the grain-filling stage significantly increased due to epoxiconazole application.Foliar application of epoxiconazole at 0.08 g/L increased grain protein content and decreased both chalky rice rate and chalkiness area ratio of fragrant rice.Epoxiconazole also substantially increased grain 2-AP content by inducing the regulation in contents of related synthetic precursors,including proline,pyrroline-5-carboxylic acid,Δ1-pyrroline and methylglyoxal.Overall,foliar application of epoxiconazole could be used for the improvement in grain yield,grain quality and 2-AP content in fragrant rice production when applied concentration at 0.08-0.32 g/L.Our findings provided the new roles of epoxiconazole in crop production.

Effects of Different Vermicompost Rates on Growth,2-Acetyl-1-Pyrroline,Photosynthesis and Antioxidant Responses of Fragrant Rice(Oryza sativa L.)Seedling

Vermicompost is an organic fertilizer contains multiple nutrient elements.However,the application of vermicompost in fragrant rice production is rarely reported.In order to study the effects of vermicompost application on fragrant rice(Oryza sativa L.)seedling performances,present study was conducted with two fragrant rice cultivars and four vermicompost rate treatments(0(CK),2.5(Wo1),5.0(Wo2)and 10.0(Wo3)g kg−1).The results showed that vermicompost treatments significantly increased dry weight of fragrant rice seedling by 8.31–32.56%compared with CK.21.10–59.13%higher net photosynthetic rates and 10.66–59.16%higher chlorophyll contents(chlorophyll a,chlorophyll b and total chlorophyll)were recorded in vermicompost treatments than CK.Application of vermicompost also significantly increased 2-acetyl-1-pyrroline(2-AP,the key compound of fragrant rice aroma)content and reduced the transcript level of gene BADH2 which related to 2-AP biosynthesis in fragrant rice seedling.Moreover,compared with CK,vermicompost treatments enhanced activities of superoxide dismutase,peroxidase,catalase by 24.42–28.66%,24.98–25.73%and 22.45–23.57%,respectively.11.54–40.53%lower malonaldehyde contents were recorded in vermicompost treatments in related to CK.In conclusion,vermicompost improved growth,increased 2-AP content and might enhance stress resistant of fragrant rice seedling.

Effects of Exogenousα-Ketoglutaric Acid on 2-Acetyl-1-Pyrroline,Yield Formation and Grain Quality Characters of Aromatic Rice

The improvement of grain quality in aromatic rice is very important for farmer to increase their income.Present study was conducted with a two-year field experiment and three aromatic rice cultivars in order to study the effects of exogenousα-ketoglutaric acid on yield formation,grain quality characters and the biosynthesis of 2-acetyl-1-pyrroline(2-AP,key component of aromatic rice’s fragrance)in aromatic rice.At heading stage,0.50 mmol L^(-1)(T1)and 1.00 mmol L^(-1)(T2)α-ketoglutaric acid solutions were overhead sprinkle to aromatic rice plants,respectively while the treatment which was overhead sprinkled with distilled water was set as control(CK).The results showed that 17.34%-33.04%and 21.39%-34.74%higher grain 2-AP contents were recorded in T1 and T2 treatments,respectively.Compared with CK,T1 and T2 treatments significantly reduced the transcript level of gene BADH2 which is related to the 2-AP biosynthesis in aromatic rice.3.86%-7.51%higher grain protein contents and 1.15%-3.37%higher head rice rates were also recorded inα-ketoglutaric acid treatments than CK.Moreover,T1 and T2 treatments remarkably decreased the chalky rice rate,chalkiness and grain amylose content.However,there was no remarkable difference in grain yield and related trails(effective panicle number,grain number per panicle,seed-setting rate and 1000-grain weight)among CK,T1 and T2 treatments.In conclusion,application of exogenousα-ketoglutaric acid enhanced 2-AP biosynthesis and improved grain quality of aromatic rice.

Enhancement of Ultrasonic Seed Treatment on Yield,Grain Quality Characters,and 2-Acetyl-1-Pyrroline Biosynthesis in Different Fragrant Rice Genotypes

Fragrant rice is popular for the good grain quality and special aroma.The present study conducted a field experiment to investigate the effects of ultrasonic seed treatment on grain yield,quality characters,physiological properties and aroma biosynthesis of different fragrant rice genotypes.The seeds of three fragrant rice genotypes were exposed to 1 min of ultrasonic vibration and then cultivated in paddy field.The results of present study showed that ultrasonic seed treatment increased grain yield of all fragrant rice genotypes but the responses of yield formation to ultrasonic were varied with different genotypes.Compared with control,ultrasonic seed treatment increased grain 2-acetyl-1-pyrroline(2-AP,the key component of fragrant rice aroma)content by 13.40%–44.88%.Ultrasonic seed treatment also reduced the crude protein contents in grains.The head rice rate,rice length,chalky rice rate,and chalkiness degree were influenced by ultrasonic for one or two fragrant rice genotypes.The activities of peroxidase and superoxide dismutase were also enhanced due to ultrasonic seed treatment.In conclusion,ultrasonic seed treatment increased grain,regulated grain aroma and quality,and improved stress resistance of fragrant rice varieties.

Comparison of 2-Acetyl-1-Pyrroline (2AP) in Rice Leaf at Different Growth Stages Using Gas Chromatography

Aromatic rice lines were examined for 2-Acetyl-1-Pyrroline (2AP) content in leaf tissue at five different growth stages (tillering, panicle initiation, 50% heading, booting, and maturity). A small plot trial with plot size of 1.42 m × 4.88 m (7 row-plots) was arranged in completely randomize design with three replications. Dry-seeded, delayed flood cultural practice was used in this study. The experiment was conducted at three locations. The average 2AP concentrations in leaf tissue at tillering stages were higher than the other four growth stages. 2AP levels were declined when rice plant reached booting. AP levels decreased slightly at heading stage and decreased significantly at maturity. There was no significant different between 2AP in leaf at 50% heading from three locations as well as the 2AP content in rice grain. Correlations between 2AP in leaf and 2AP in grain were significantly in all five growth stages. The highest correlation coefficient was found between 2AP in leaf at booting and grain (r = 0.811**) and lowest was in the leaf at harvest (r = 0.564**). Results indicated that 2AP could be determined in leaf tissue at early growth stage.

稻米香味物质2-乙酰-1-吡咯啉的形成及其影响因素

稻米香味的形成是一个复杂而多变的生理过程,遗传和环境变化对香味的形成都具有重要影响。香味的有无主要受香味基因BADH2控制,气候条件、土壤质地和栽培措施也会直接影响稻米香味物质的合成。为进一步加快香稻培育进程,提升香稻稻米品质,本文综述了近年来国内外有关稻米香味物质形成的遗传和生理机制以及环境因素对稻米香味形成的研究进展,重点总结了外界光照、水分、温度、肥料、矿质元素等对稻米香味物质合成的影响,并指出了今后研究的方向,以期为提高我国香稻栽培技术,提升香稻稻米品质提供参考。

新型香稻渝恢2103香味分子遗传特性分析

香味是优良稻米品质的重要衡量标准之一,2-乙酰-1-吡咯啉(2AP)是最主要的香味物质,然而2AP生物合成机理至今仍未确凿。本研究筛选了与2AP生物合成密切相关的甜菜碱脱氢酶2基因(Badh2)在30份水稻材料中的3种突变类型,从中发现1份新的香稻材料渝恢2103,该材料Badh2基因序列编码区无突变,遗传分析显示渝恢2103与badh2-E7突变型香稻宜香1B香味基因不等位,与非香稻杂交F2香与非香分离比接近9∶7,与香稻杂交F2香与非香分离比接近7∶9,表明渝恢2103的香味受多基因控制。进一步利用实时荧光定量PCR技术(qRT-PCR)比较了与2AP生物合成相关基因在日本晴、渝恢2103和宜香1B的表达情况,结果显示,Badh2基因在日本晴和渝恢2103中表达差异不大,但在宜香1B中表达量异常高;多数脯氨酸与谷氨酸代谢途径相关基因在宜香1B中的表达水平显著高于日本晴和渝恢2103;推测宜香1B的2AP合成同时受Badh2基因以及脯氨酸与谷氨酸代谢途径相关基因的影响;渝恢2103香味形成可能与这些基因无必然联系。渝恢2103特殊的遗传特性可能为水稻香味形成机理研究提供新的突破点。

肥料类型及施用方式对香稻香气2-乙酰-1-吡咯啉含量的影响

为了探明肥料类型及施用方式对香稻香气2-乙酰-1-吡咯啉的影响及其生理基础的影响,以大面积推广应用的香稻品种玉香油占为研究材料开展大田试验。试验设置深施肥和常规手撒施肥2种施肥方式处理,施用碳酸氢铵+过磷酸钙+氯化钾处理和复合肥处理2种肥料类型处理,各肥料处理均含有等质量的氮磷钾养分。研究了香稻成熟期籽粒香气2AP的含量,分蘖期、孕穗期、扬花期和成熟期香稻叶片的脯氨酸含量、脯氨酸脱氢酶活性、吡咯啉-5-羧酸合成酶活性、鸟氨酸转氨酶活性和吡咯啉-5-羧酸含量。结果表明,肥料类型、肥料施用方式和两者互作均显著影响香稻籽粒香气2AP含量,其中常规手撒施用碳酸氢铵+过磷酸钙+氯化钾肥料处理利于香稻籽粒积累显著较高的香气2AP含量,达59.86 ng/g;香稻籽粒香气2AP含量和成熟期叶片的吡咯啉-5-羧酸含量呈显著正相关关系(r=0.957 1*)。该研究可为制定香稻浓香施肥技术提供理论参考依据。

香稻香气特征化合物2–乙酰基–1–吡咯啉测定方法的建立

建立了一种利用顶空固相微萃取–气相色谱法（HS–SPME–GC）检测香稻香味特征化合物2–乙酰基–1–吡咯啉（2–acetyl–1–Pyrroline,2–AP）的方法。在合成2–AP标准物质的基础上,通过正交试验得到最佳萃取条件为萃取温度60℃、萃取时间45 min。在最佳萃取条件下,该检测方法的线性范围为0.5~4.0μg/g,相对标准偏差≤8.54%,回收率为96.3%~103.5%,检出限为0.045 5μg/g,定量限为0.152μg/g。利用该方法测定6种不同香稻大米2–AP的含量,结果表明,该方法能有效地检测香米的2–AP含量。

香稻品种2-乙酰-1-吡咯啉多样性及籽粒分布特征的研究

采用田间试验方法研究同一地点和时间种植、同一栽培条件下2-乙酰-1-吡咯啉在11个香稻品种间差异性及籽粒分布特征。结果表明,桂香丝糯、中健2号、清香米、泰国香稻1号R207和Texmati等5个品种含有2-乙酰-1-吡咯啉,精米与糠中的2-乙酰-1-吡咯啉含量在上述5个品种间存在显著差异(P<0.05),其中Texmati精米和糠中2-乙酰-1-吡咯啉的含量最高,分别为0.6765和0.1738mg/kg,桂香丝糯精米和糠中2-乙酰-1-吡咯啉的含量最低,分别为0.0452和0.0173mg/kg。2-乙酰-1-吡咯啉在香稻籽粒中分布特征为90%以上分布于精米,不足10%分布于糠,稻壳中未见2-乙酰-1-吡咯啉分布。

香米特征香味成分2-乙酰基-1-吡咯啉提取及检测方法研究进展

2-乙酰基-1-吡咯啉(2-AP)是目前已经探明的香米特征性香味成分,其含量的高低直接影响香米的品质与价格,但由于该成分在天然香米中的含量通常低至微克每千克,在采用现代分析检测方法测定时,还可能受到样品基质干扰、色谱分离共流出等问题的影响,因此,如何从稻米样品中高效提取该化合物并结合适当的检测方法进行精确测定具有一定的挑战性。该文综述了对稻米中2-AP进行测定时常用的样品前处理方法及仪器分析方法。样品前处理方法主要有蒸馏法、萃取法及顶空富集法等;仪器分析方法主要包括气相色谱及气相色谱-质谱法、气相色谱-嗅觉测试法,以及最新被开发出来的衍生化-高效液相色谱-串联质谱法。该综述将为食品贸易领域香米中2-AP的测定、农业领域香米的遗传育种及水肥管理、食品加工领域米香类稳定风味成分的开发等提供参考依据。

大米中2-乙酰基-1-吡咯啉检测技术及形成机制研究进展

2-乙酰基-1-吡咯啉是大米中的特征香气成分,其含量高低直接影响大米品质和价格。因此,开展大米中2-乙酰基-1-吡咯啉精准检测方法研究,对于明确呈香机理、提升大米品质及促进进出口贸易都具有重要意义。综述了当前主要的前处理技术和基于GC、GC-MS、GC-O-MS、GC-TOF-MS、GC×GC-TOF-MS、HPLC-MS/MS等仪器检测2-乙酰基-1-吡咯啉的研究进展,分析了各种方法的优缺点,以及2-乙酰基-1-吡咯啉形成机制的研究进展,以期实现大米中2-乙酰基-1-吡咯啉的精准检测及大米品质提升提供借鉴。

γ-氨基丁酸和2-乙酰-1-吡咯啉代谢通路在水稻籽粒发育过程中的变化分析

【目的】结合代谢组和转录组技术,分析香稻和非香稻不同发育时期籽粒中γ-氨基丁酸(GABA)和2-乙酰-1-吡咯啉(2AP)代谢通路主要化合物和基因的动态变化,为高含量GABA和2AP的水稻育种提供理论依据。【方法】本研究收集优质香籼稻美香占2号和优质籼稻黄华占花后8 d(乳熟期)、花后15 d(蜡熟期)、30 d(完熟期)的脱壳籽粒和花后40 d收获的籽粒的精米,对各样本采用LC-MS/MS和RNA-seq技术进行代谢产物检测和转录表达分析,分析两个水稻品种GABA和2AP代谢通路相关化合物的空间分布以及含量变化特征。【结果】代谢组数据PCA分析表明两个水稻品种乳熟期籽粒和花后40 d收获的籽粒的精米明显分离,蜡熟期和完熟期籽粒分离不明显。代谢组共鉴定出623种代谢物,四个时期差异代谢物合计161个。GABA和2AP通路中9个代谢物被检出,两个水稻品种的相关代谢物变化趋势相似,其中腐胺主要定位于籽粒的糊粉层,而亚精胺在内胚乳特异累积。转录组共检出3.3万个左右基因,其中检出GABA通路18个酶中的14个酶对应的基因有较高的表达水平,除Badh2外其他基因均不是差异表达基因,同一基因在两份水稻材料中的表达水平和变化趋势相似,GAD1、GABA-T1、DAO4和PAO4在相应同源基因中表达水平较高。黄华占籽粒中L-精氨酸和腐胺的代谢在蜡熟期已基本结束,美香占2号在籽粒整个发育期具有持续较强的多胺降解水平。美香占2号籽粒具有合成2AP的物质基础,多胺降解途径和谷氨酸-脯氨酸转化通路同时有助于2AP的积累。【结论】水稻籽粒GABA代谢以降解为主,GABA-T1和GABA-T2共同调控GABA的降解。GABA相关代谢物主要集中在糊粉层,黄华占籽粒中代谢物分布和基因表达特征与美香占2号相似,美香占2号籽粒成熟过程中具有持续较高的线粒体活性。

利用CRISPR/Cas9技术编辑BADH2-1/BADH2-2创制香米味道玉米新种质

【目的】香味是作物的重要食味品质之一。2-乙酰-1-吡咯啉(2-acetyl-1-pyrroline,2-AP)是主要香味物。BADH2是控制水稻等作物香味性状的关键基因,敲除该基因可以产生香味稻米。利用CRISPR/Cas9基因编辑技术在北京市农林科学院自育的玉米骨干亲本京724上敲除BADH2同源基因,以期获得有香米味道的玉米新种质材料。【方法】利用Ensembl数据库在线BLAST工具,将水稻OsBADH2蛋白序列在拟南芥、水稻和玉米蛋白序列数据库中进行序列比对,获得上述3个物种的BADH基因家族成员,并利用UniProt蛋白数据库中的结构域信息进行验证。进一步使用MEGA软件进行系统进化分析,获得玉米BADH2同源基因作为候选编辑靶标。基于CRISPR/Cas9基因编辑技术的原理,在候选基因的外显子处设计特异性靶点,并构建入CRISPR/Cas9基因编辑载体。再以玉米自交系京724为受体,利用农杆菌介导的遗传转化方法,通过磷酸甘露糖异构酶基因(phosphomannose isomerase,PMI)抗性筛选获得阳性转基因植株。转基因株系经测序明确其在靶基因中产生的突变类型。利用气相色谱质谱联用仪(gas chromatography-mass spectrometry,GC-MS)检测基因编辑株系T1籽粒中香米主要香味物质2-AP的含量,以确认京724在基因编辑前后2-AP含量的变化。【结果】系统进化分析发现,玉米中存在2个BADH2同源基因,分别命名为ZmBADH2-1和ZmBADH2-2。ZmBADH2-1位于第4染色体,ZmBADH2-2位于第1染色体。2个基因均包含15个外显子和14个内含子,第4外显子间的核苷酸序列高度同源。在2个基因的第4外显子区域设计靶点并构建入CRISPR/Cas9基因编辑载体,通过遗传转化获得28株转基因株系。PCR扩增及测序分析结果显示,其中10株材料的2个ZmBADH2s在靶点区域均发生突变,突变基因型包括双等位突变和多等位突变,突变类型为不同数量的碱基缺失和插入。质谱检测结果显示玉米ZmBADH2双基因突变体籽粒中存在与香稻同样成分的2-AP。随机选取的4个T_(1)代基因编辑株系籽粒中,2-AP平均含量分别为438.29、404.63、348.65和161.82μg·kg-1,而未经过编辑的京724中未检测到2-AP。【结论】利用CRISPR/Cas9技术对玉米ZmBADH2-1和ZmBADH2-2同时进行定点敲除,创制出籽粒中具有香米味道的玉米骨干亲本新种质材料。

香稻Badh2基因单倍型及香气成分2-乙酰-1-吡咯啉代谢通路的研究进展

香米因其具有独特的香味,长期以来深受国际市场青睐。我国香稻种植历史悠久,香稻资源丰富,但缺乏高端优质香米品牌。当前消费者对优质香米的需求不断加大,进而加快了我国优质香稻的育种进程。稻米香味主要由编码甜菜碱醛脱氢酶基因Badh2控制,该基因功能突变能够导致稻米香气成分2-乙酰-1-吡咯啉(2AP)的积累。随着水稻功能基因组学的发展,香稻资源中Badh2基因单倍型得以深入挖掘,2AP生物合成通路被初步解析,它们将进一步加快香稻育种的进程。综述了目前在香稻资源中发现的19种Badh2基因单倍型,重点分析不同类型单倍型的地域分布特性,比较不同单倍型之间的2AP含量;同时阐述2AP生物合成通路涉及的谷氨酸-脯氨酸代谢途径和多胺代谢途径研究现状,总结香稻籽粒中2AP的分布特征,并对目前两种籽粒2AP积累的理论机制进行比较分析,提出利用多组学手段从籽粒动态发育过程研究2AP生物合成通路变化特性的策略,以期为香稻资源基因利用和籽粒2AP特异富集香稻品种的培育提供参考。

利用CRISPR-Cas9技术编辑Badh2基因创制优质香型籼稻三系不育系

稻米香味是水稻的重要食味品质之一,其主要受第8染色体上编码甜菜碱脱氢酶基因Badh2控制,该基因突变可导致香味物质2-乙酰-1-吡咯啉(2-AP)的含量增加从而促进香味的产生。本研究以三明市农业科学研究院自主选育的优质籼型杂交稻保持系明太B为受体,利用CRISPR-Cas9技术对其Badh2基因进行编辑和敲除。获得2个T0代转基因纯合突变体植株并对其衍生的48个T1代单株进行鉴定和分析,获得1个不含转基因载体骨架且在第2外显子插入单个碱基T的纯合突变体株系明太B-badh2。利用半定量PCR和qRT-PCR技术以及气相色谱质谱联用仪(GC-MS)检测Badh2基因相对表达量和2-AP含量;同时采用农业行业标准(NY/T1433-2014)推荐的48对水稻SSR引物进行指纹图谱分析。结果表明,该株系Badh2基因RNA表达水平显著下调;籽粒中香味物质2-AP的含量显著增加;指纹图谱分析发现,仅1对引物Rm571在野生型和突变体之间鉴定到等位变异,两组材料遗传差异较小。此外,本研究还对野生型和突变体T2代植株表型性状、稻米蒸煮食味品质和外观品质指标进行了考察和测定分析。结果表明,所有指标在两组材料间均无显著差异。进一步采用测交和回交转育方法并结合Badh2位点测序分析,成功选育获得了其对应的纯合香型三系不育系明太A-badh2。通过与恢复系明恢703、明恢3009测配,其组合产量与国家审定品种明太优703、明太优3009相近且表现出较强的超标优势。此外,通过与香型恢复系明恢1831测配后发现其组合籽粒中香味物质2-AP含量极显著高于对照组合明太A/明恢1831。因此,利用CRISPR-Cas9基因编辑技术,可对水稻香味基因Badh2进行精准定向编辑和敲除,实现对水稻香味性状的改良,为创制香型籼稻不育系提供理论指导,从而加快香型杂交稻育种进程。

比较代谢组和转录组学揭示“19香”水稻香气物质组成及其形成途径

基于气象色谱-质谱联用(Gas Chromatography-Mass Spectrometry,GC-MS)对广东省龙川市和连山市种植的“19香”乳熟期籽粒胚乳的挥发性代谢物进行代谢组学分析,结合转录组技术挖掘香气物质形成的相关基因,探讨产地对香稻香气物质形成的影响。通过主成分分析(Principal Component Analysis,PCA)、偏最小二乘法判别分析(Partial Least Squares Discriminant Analysis,PLS-DA)对原始数据进行多元统计分析。结果表明,“19香”异地种植后其挥发性物质含量发生变化,其中有14个香气物质的相对含量达显著差异。通过KEGG富集分析发现次级代谢物的生物合成对香气物质含量具有显著影响。在转录水平上以连山组为对照,龙川组样本有56个显著上调表达基因和89个下调表达基因,且主要参与有机物的分解代谢和多糖代谢等生物进程,差异表达基因发挥着糖基键的水解酶活性和氧化还原酶活性等分子功能,此外类胡萝卜素合成途径为显著富集通路。综上所述,不同种植环境对香稻的香气物质含量影响较大,其中乳熟期糖类代谢活动、胁迫应答相关机制可能是种植环境中调控香气物质含量的关键因子。

苗期喷施茉莉酸甲酯对香稻品种象牙香占香气的影响

茉莉酸甲酯能够提高植物的抗逆境胁迫能力,但在对香稻香气物质2-乙酰基-1-吡咯啉(2-AP)的影响研究还鲜有报道。以香稻品种象牙香占为试验材料,设置茉莉酸甲酯3种喷施浓度(0.25、0.50和1.00 mmol/L),研究苗期喷施对香稻幼苗2-AP合成与累积、香气前体物质含量、香气相关酶活性、香气相关基因表达量调控、抗性酶活性变化和秧苗素质的影响。结果表明,与对照(CK)相比,喷施适当的茉莉酸甲酯浓度(0.50~1.00 mmol/L),香稻幼苗2-AP含量增加,香气合成前体物质1-吡咯啉、丙酮醛和1-吡咯啉-5-羧酸(P5C)含量提高,二胺氧化酶2基因(DAO4)、鸟氨酸转氨酶基因(OAT)和脯氨酸脱氢酶基因(ProDH)表达量提升。茉莉酸甲酯处理还提高了香稻秧苗素质,增加了抗性酶活性,提高了抗氧化能力。适度的茉莉酸甲酯能提高香稻2-AP含量和秧苗素质,归因于相关酶活性和基因表达量的提高。

气相色谱-质谱技术分析香稻特征化合物2-乙酰基吡咯啉

建立了气相色谱-质谱技术(GC-MS)分析香稻特征化合物2-乙酰基吡咯啉的方法。在香稻中加入内标物2,4,6-三甲基吡啶,密闭,经无水乙醇-二氯甲烷(体积比为1∶1)在80℃水浴中提取3h。考察了温度和时间对2-乙酰基吡咯啉提取的影响。提取液经毛细管气相色谱柱HP-5MS(30m×0.25mm×0.25μm)分离,全扫描监测模式GC-MS测定。以内标物计,方法的平均回收率为82.57%,相对标准偏差为5.09%,最低检出限为0.01mg/kg。将该方法应用于11份香稻育种材料中2-乙酰基吡咯啉的测定,结果表明,清香米、泰香R207、Texmati、桂香丝糯和中健2号等5个品种中含有2-乙酰基吡咯啉,含量分别为0.097、0.098、0.699、0.045和0.047mg/kg。该方法样品处理简单、快速、灵敏、样本和试剂消耗少,尤其适合于通过测定2-乙酰基吡咯啉含量进行香稻育种前的大批量品种筛选。