Modified Atmospheric Packaging and Its Effect on Postharvest Cannabis Quality

Cannabis sativa L. is used as fiber, food, and medicine in several countries. Though it is illegal for recreational use in most of the world, there are some countries that have legalized production and sale. There is a lot of research on production of cannabis, but less so on storage technologies. Cannabis contains several high value compounds, such as cannabinoids and terpenoids, that are susceptible to degradation via light, temperature, and oxygen. Several studies have explored temperature and light, and industry has adjusted accordingly. However, less is known about oxygen-induced degradation. Biochemical studies have demonstrated oxidative degradation of high value compounds, and many producers use some form of modified atmospheric packaging (MAP) for storage. However, the efficacy of MAP is unclear. The objective of this paper is to review our current understanding of MAP in postharvest cannabis storage and identify avenues where additional research is needed.

大麻(Cannabis sativa L.)内生细菌的分离与鉴定

为分离与鉴定大麻(Cannabis sativa L.)雌株及雄株3个部位的叶片(从植株茎基向茎尖方向第二轮叶,第五轮叶,雄花周围叶)和花的内生细菌,本研究采用微生物培养和分子生物学鉴定方法,探讨其种类及其分布特征。结果显示,从大麻雄株中分离到6属35株内生细菌,大麻雌株中分离到11属39株内生细菌。内生细菌种属在大麻雌雄植株不同部位的叶和花中的分布存在显著差异,大麻雌株与雄株的内生细菌种类只共有葡萄球菌属(Staphylococcus sp.)、芽孢杆菌属(Bacillus sp.)和马赛菌属(Massilia sp.),其余均不共有。大麻雄株3个部位叶片中共有阿尔莱特葡萄球菌和溶血葡萄球菌而雄花中不存在;在雄花中分离到地衣芽孢杆菌、木糖葡萄球菌和贝莱斯芽孢杆菌而叶中不存在。大麻雌株3个部位叶片共有贝莱斯芽孢杆菌和特基拉芽孢杆菌而雌花中不存在;在雌花中分离到粪短波单胞菌、谷粒申氏菌、农杆菌、香料葡萄球菌、缺陷短波单胞菌、地衣芽胞杆菌和弗留明拜叶林克氏菌而叶中不存在。结果表明大麻内生细菌在不同植株、叶和花中的多样性以及不同内生细菌的分布差异。本研究结果为进一步研究大麻内生细菌的多样性提供科学依据,同时为未来大麻内生菌功能性促生菌种的筛选提供基础。

The auxin transporter OsAUX1 regulates tillering in rice(Oryza sativa)

Tillering is an important agronomic trait of rice(Oryza sativa)that affects the number of effective panicles,thereby affecting yields.The phytohormone auxin plays a key role in tillering.Here we identified the high tillering and semi-dwarf 1(htsd1)mutant with auxin-deficiency root characteristics,such as shortened lateral roots,reduced lateral root density,and enlarged root angles.htsd1 showed reduced sensitivity to auxin,but the external application of indole-3-acetic acid(IAA)inhibited its tillering.We identified the mutated gene in htsd1 as AUXIN1(OsAUX1,LOC_Os01g63770),which encodes an auxin influx transporter.The promoter sequence of OsAUX1 contains many SQUAMOSA PROMOTER BINDING PROTEIN-LIKE(SPL)binding sites,and we demonstrated that SPL7 binds to the OsAUX1 promoter.TEOSINTE BRANCHED1(OsTB1),a key gene that negatively regulates tillering,was significantly downregulated in htsd1.Tillering was enhanced in the OsTB1 knockout mutant,and the external application of IAA inhibited tiller elongation in this mutant.Overexpressing OsTB1 restored the multi-tiller phenotype of htsd1.These results suggest that SPL7 directly binds to the OsAUX1 promoter and regulates tillering in rice by altering OsTB1 expression to modulate auxin signaling.

Irrigation regimes modulate non-structural carbohydrate remobilization and improve grain filling in rice(Oryza sativa L.)by regulating starch metabolism

Recently developed ‘super’ rice cultivars with greater yield potentials often suffer from the problem of poor grain filling, especially in inferior spikelets. Here, we studied the activities of enzymes related to starch metabolism in rice stems and grains, and the microstructures related to carbohydrate accumulation and transportation to investigate the effects of different water regimes on grain filling. Two ‘super’ rice cultivars were grown under two irrigation regimes of well-watered(WW) and alternate wetting and moderate soil drying(AWMD). Compared with the WW treatment,the activities of ADP glucose pyrophosphorylase(AGPase), starch synthase(StSase) and starch branching enzyme(SBE), and the accumulation of non-structural carbohydrates(NSCs) in the stems before heading were significantly improved, and more starch granules were stored in the stems in the AWMD treatment. After heading, the activities of α-amylase, β-amylase, sucrose phosphate synthase(SPS) and sucrose synthase in the synthetic direction(SSs)were increased in the stems to promote the remobilization of NSCs for grain filling under AWMD. During grain filling, the enzymatic activities of sucrose synthase in the cleavage direction(SSc), AGPase, StSase and SBE in the inferior spikelets were increased, which promoted grain filling, especially for the inferior spikelets under AWMD.However, there were no significant differences in vascular microstructures. The grain yield and grain weight could be improved by 13.1 and 7.5%, respectively, by optimizing of the irrigation regime. We concluded that the low activities of key enzymes in carbon metabolism is the key limitation for the poor grain filling, as opposed to the vascular microstructures, and AWMD can increase the amount of NSC accumulation in the stems before heading, improve the utilization rate of NSCs after heading, and increase the grain filling, especially in the inferior spikelets, by altering the activities of key enzymes in carbon metabolism.

基于水稻大粒染色体片段代换系Z29的鉴定及QTL定位

千粒质量作为水稻产量的三要素之一,对产量的影响较大,其中千粒质量主要受水稻粒型的影响,因此挖掘新的粒型基因在生产中具有重要的意义.研究选育到1个以日本晴为受体亲本、自育优良籼稻恢复系R225为供体亲本的水稻染色体片段代换系(CSSL)Z29. Z29含有来自R225的10个代换片段,平均代换长度2.90 Mb. Z29粒长和粒宽均极显著增加,表现为大粒表型,且其籽粒变大是由颖壳细胞数量极显著增多、增大引起.利用日本晴与Z29杂交构建的次级F2群体进行QTL定位,共检测到8个粒型相关QTL.进一步利用MAS法在F3群体中选育出14个次级染色体片段代换系,包括4个单片段代换系、 5个双片段代换系、 2个三片段代换系和3个四片段代换系.结果可为目的粒型相关QTL克隆和分子机制解析奠定基础,为分子设计育种提供资源.

Endophytic fungi harbored in Cannabis sativa L.:diversity and potential as biocontrol agents against host plant-specific phytopathogens

The objective of the present work was isolation,phylogenetic characterization,and assessment of biocontrol potential of endophytic fungi harbored in various tissues(leaves,twigs,and apical and lateral buds)of the medicinal plant,Cannabis sativa L.A total of 30 different fungal endophytes were isolated from all the plant tissues which were authenticated by molecular identification based on rDNA ITS sequence analysis(ITS1,5.8S and ITS2 regions).The Menhinick’s index revealed that the buds were immensely rich in fungal species,and Camargo’s index showed the highest tissue-specific fungal dominance for the twigs.The most dominant species was Penicillium copticola that could be isolated from the twigs,leaves,and apical and lateral buds.A detailed calculation of Fisher’s log series index,Shannon diversity index,Simpson’s index,Simpson’s diversity index,and Margalef’s richness revealed moderate overall biodiversity of C.sativa endophytes distributed among its tissues.The fungal endophytes were challenged by two host phytopathogens,Botrytis cinerea and Trichothecium roseum,devising a dual culture antagonistic assay on five different media.We observed 11 distinct types of pathogen inhibition encompassing a variable degree of antagonism(%)on changing the media.This revealed the potential chemodiversity of the isolated fungal endophytes not only as promising resources of biocontrol agents against the known and emerging phytopathogens of Cannabis plants,but also as sustainable resources of biologically active and defensive secondary metabolites.

A Comparative Study on Hemp(Cannabis sativa)Essential Oil Extraction Using Traditional and Advanced Techniques

A comparative study of Cannabis sativa(Hemp)essential constituents obtained by using Supercritical Fluid Extraction(SCFE),Steam Distillation(SD)and Hydrodistillation(HD)is presented here.The optimized extraction temperatures were 130,110and 50℃for hydrodistillation,steam distillation and supercritical fluid extraction respectively.The essential oil of C.sativa was analyzed by using Gas chromatography mass spectrometry(GC-MS).A total of 33,30and 31components have been identified in HD,SD and SCFE respectively.Yield of essential oil using SCFE(0.039%)was more than HD(0.025%)and SD(0.035%)extraction respectively.The main component of sesquiterpenes obtained by hydrodistillation at 130℃with their percentages included caryophyllene(40.58%),trans-α-bergamotene(5.41%),humulene(10.97%),cis-β-farnesene(8.53%)and monoterpenes includedα-pinene(2.13%),d-limonene(6.46%),p-cymol(0.65%)and cineole(2.58%)respectively.The main component of sesquiterpenes obtained by SD steam distillation at110℃including caryophyllene(38.60%)trans-α-bergamotene(4.22%),humulene(10.26%),cis-β-farnesene(6.67%)and monoterpenes includedα-pinene(3.21%),d-limonene(7.07%),p-cymol(2.59%)and cineole(3.88%)whereas the more percentages of major components were obtained by SCFE at 50℃included caryophyllene(44.31%),trans-α-bergamotene(6.79%),humulene(11.97%)cis-β-farnesene(9.71%)and monoterpenes includedα-pinene(0.45%),d-limonene(2.13%)p-cymol(0.19%)and cineole(1.38%)respectively.We found yield/efficiency,chemical composition,quality of the essential oils by supercritical fluid extraction superior in terms of modern,green,saving energy and a rapid approach as compared to traditional techniques.

镉胁迫对线麻(CannabisSativaL.)富集及光合特性的影响

选择广泛栽培于西北地区的线麻(Cannabis Sativa L.)进行盆栽实验,研究了Cd胁迫对线麻富集和光合特性的影响.结果显示,线麻根、茎、叶中Cd最大富集量分别达到503.40、350.63、77.90 mg·kg^(-1).随着土壤Cd浓度的增加,线麻根、茎、叶的富集量逐渐增加,且表现为:根部>茎部>叶部.当Cd≥150 mg·kg^(-1)时,Cd从根部向地上部分的转运开始加强,且地上部分组织中主要集中在茎部.线麻株高、比叶重和生物量随Cd浓度的增加先升高后下降.当Cd浓度≥150 mg·kg^(-1)时,株高、比叶重和地下部分生物量明显低于对照(P<0.05),而地上部分生物量不存在显著性差异(P>0.05).当Cd浓度为50 mg·kg^(-1)时线麻叶绿素含量、类胡萝卜素含量、净光合速率、蒸腾速率、气孔导度、最大净光合速率和光饱和点达到最大值,光补偿点随Cd浓度的增加而持续增加.结果表明,线麻具有较高的Cd富集能力,可以作为潜在的Cd污染土壤修复栽培作物,但野外修复中如何提高线麻的生物量有待于进一步研究.

DROUGHT-INDUCED UNKNOWN PROTEIN 1 positively modulates drought tolerance in cultivated alfalfa(Medicago sativa L.)

Alfalfa is the most widely cultivated perennial legume forage crop worldwide.Drought is one of the major environmental factors influencing alfalfa productivity.However,the molecular mechanisms underlying alfalfa responses to drought stress are still largely unknown.This study identified a drought-inducible gene of unknown function,designated as Medicago sativa DROUGHT-INDUCED UNKNOWN PROTEIN 1(MsDIUP1).MsDIUP1 was localized to the nucleus,chloroplast,and plasma membranes.Overexpression of MsDIUP1 in Arabidopsis resulted in increased tolerance to drought,with higher seed germination,root length,fresh weight,and survival rate than in wild-type(WT)plants.Consistently,analysis of MsDIUP1 over-expression(OE)alfalfa plants revealed that MsDIUP1 also increased tolerance to drought stress,accompanied by physiological changes including reduced malondialdehyde(MDA)content and increased osmoprotectants accumulation(free proline and soluble sugar),relative to the WT.In contrast,disruption of MsDIUP1 expression by RNA interference(RNAi)in alfalfa resulted in a droughthypersensitive phenotype,with a lower chlorophyll content,higher MDA content,and less osmoprotectants accumulation than that of the WT.Transcript profiling of alfalfa WT,OE,and RNAi plants during drought stress showed differential responses for genes involved in stress signaling,antioxidant defense,and osmotic adjustment.Taken together,these results reveal a positive role for MsDIUP1 in regulating drought tolerance.

Cannabinoids Production by Hairy Root Cultures of <i>Cannabis sativa</i>L.

Tetrahydrocannabinol (THC) derivatives are used clinically as analgesic, anti-inflammatory, appetite stimulant, anti-emetic and anti-tumor cannabinoids. THC and its related compounds are at present obtained by extraction from intact Cannabis plants or chemical synthesis, but plant cell cultures may be an alternative source of production. In the present study, hairy root cultures of C. sativa (Cannabaceae) were induced by incubation of aseptically grown callus culture with solid B5 medium supplemented with 4 mg/l naphthaleneacetic acid in darkness at 25°C. Hairy root growth profiles in shake flask, increased periodically during 35 days of growth cycle. The cannabinoid contents produced in minor levels and remained below 2.0 μg/g dry weight. The contents of can-nabinoid were analyzed by liquid chromatography and confirmed by mass spectrometry.

Conventional Breeding and Molecular Markers for Blast Disease Resistance in Rice (Oryza sativa L.)

Monogenic lines,which carried 23 genes for blast resistance were tested and used donors to transfer resistance genes by crossing method.The results under blast nursery revealed that 9 genes from 23 genes were susceptible to highly susceptible under the three locations(Sakha,Gemmeza,and Zarzoura in Egypt);Pia,Pik,Pik-p,Piz-t,Pita,Pi b,Pi,Pi 19 and Pi 20.While,the genes Pii,Pik-s,Pik-h,Pi z,Piz-5,Pi sh,Pi 3,Pi 1,Pi 5,Pi 7,Pi 9,Pi 12,Pikm and Pita-2 were highly resistant at the same locations.Clustering analysis confirmed the results,which divided into two groups;the first one included all the susceptible genes,while the second one included the resistance genes.In the greenhouse test,the reaction pattern of five races produced 100%resistance under artificial inoculation with eight genes showing complete resistance to all isolates.The completely resistant genes:Pii,Pik-s,Piz,Piz-5(=bi2)(t),Pita(=Pi4)(t),Pita,Pi b and Pi1 as well as clustering analysis confirmed the results.In the F1 crosses,the results showed all the 25 crosses were resistant for leaf blast disease under field conditions.While,the results in F2 population showed seven crosses with segregation ratio of 15(R):1(S),two cross gave segregated ratio of 3 R:1 S and one gave 13:3.For the identification of blast resistance genes in the parental lines,the marker K3959,linked to Pik-s gene and the variety IRBLKS-F5 carry this gene,which was from the monogenic line.The results showed that four genotypes;Sakha 105,Sakha 103,Sakha 106 and IRBLKS-F5 were carrying Pik-s gene,while was absent in the Sakha 101,Sakha 104,IRBL5-M,IRBL9-W,IRBLTACP1 and IRBL9-W(R)genotypes.As for Pi 5 gene,the results showed that it was present in Sakha 103 and Sakha 104 varieties and absent in the rest of the genotypes.In addition,Pita-Pita-2 gene was found in the three Egyptian genotypes(Sakha 105,Sakha 101 and Sakha 104)plus IRBLTACP1 monogenetic.In F2 generation,six populations were used to study the inheritance of blast resistance and specific primers to confirm the ratio and identify the resistance genes.However,the ratios in molecular markers were the same of the ratio under field evaluation in the most population studies.These findings would facilitate in breeding programs for gene pyramiding and gene accumulation to produce durable resistance for blast using those genotypes.

全基因组关联分析定位水稻分蘖角度QTL

【目的】水稻分蘖角度是影响水稻产量的关键农艺性状,挖掘水稻分蘖角度QTL(基因)及其优势单倍型,有助于构建水稻理想株型。【方法】以333份来自水稻3K资源的核心种质为研究材料,于2020年和2022年分别在湖南农业大学耘园基地和春华基地种植,在抽穗期测量分蘖角度,结合基因型,利用TASSEL 5.2的MLM模型进行全基因组关联分析。【结果】共检测到6个分蘖角度QTL位点,分布在水稻2、5、6、9和12号染色体上,分别命名为qTA2、qTA5、qTA6.1、qTA6.2、qTA9和qTA12,这些QTL的表型贡献率为6.23%~16.22%。除了qTA9与分蘖角度主效QTL TAC1共定位外,其余5个QTL均为新的位点。进一步对5个QTL位点进行候选基因分析,初步筛选到qTA2和qTA6.1的候选基因别为Os02g0817900和Os06g0682800,候选基因Os02g0817900编码水稻细胞色素P450家族蛋白,候选基因Os06g0682800编码锌指结构域蛋白。【结论】本研究挖掘到新的水稻分蘖角度相关位点并对候选基因进行了分析,为分蘖角度新QTL(基因)的克隆以及分蘖角度的遗传改良提供参考。

A Farmer’s Approach to Detecting Photoperiod Sensitivity in Rice (Oryza sativa ssp. indica) Landraces

Most indigenous rice landraces are sensitive to photoperiod during short day seasons,and this sensitivity is more pronounced in indica than in japonica landraces.Attempts to identify photoperiod sensitive(PPS)cultivars based on the life history stages of the rice plant,and several models and indices based on phenology and day length have not been precise,and in some cases yield counterfactual inferences.Following the empirical method of traditional Asian rice farmers,the author has developed a robust index,based on the sowing and flowering dates of a large number of landraces grown in different seasons from 2020 to 2023,to contradistinguish PPS from photoperiod insensitive cultivars.Unlike other indices and models of photoperiod sensitivity,the index does not require the presumed duration of different life history stages of the rice plant but relies only on the flowering dates and the number of days till flowering of a rice cultivar sown on different dates to consistently identify photoperiod sensitive cultivars.

The response of roots and the rhizosphere environment to integrative cultivation practices in paddy rice

Integrative cultivation practices(ICPs)are essential for enhancing cereal yield and resource use efficiency.However,the effects of ICP on the rhizosphere environment and roots of paddy rice are still poorly understood.In this study,four rice varieties were produced in the field.Each variety was treated with six different cultivation techniques,including zero nitrogen application(0 N),local farmers’practice(LFP),nitrogen reduction(NR),and three progressive ICP techniques comprised of enhanced fertilizer N practice and increased plant density(ICP1),a treatment similar to ICP1 but with alternate wetting and moderate drying instead of continuous flooding(ICP2),and the same practices as ICP2 with the application of organic fertilizer(ICP3).The ICPs had greater grain production and nitrogen use efficiency than the other three methods.Root length,dry weight,root diameter,activity of root oxidation,root bleeding rate,zeatin and zeatin riboside compositions,and total organic acids in root exudates were elevated with the introduction of the successive cultivation practices.ICPs enhanced nitrate nitrogen,the activities of urease and invertase,and the diversity of microbes(bacteria)in rhizosphere and non-rhizosphere soil,while reducing the ammonium nitrogen content.The nutrient contents(ammonium nitrogen,total nitrogen,total potassium,total phosphorus,nitrate,and available phosphorus)and urease activity in rhizosphere soil were reduced in all treatments in comparison with the non-rhizosphere soil,but the invertase activity and bacterial diversity were greater.The main root morphology and physiology,and the ammonium nitrogen contents in rhizosphere soil at the primary stages were closely correlated with grain yield and internal nitrogen use efficiency.These findings suggest that the coordinated enhancement of the root system and the environment of the rhizosphere under integrative cultivation approaches may lead to higher rice production.

Determination of Optimal Hormone and Mineral Salts Levels in Tissue Culture Media for Callus Induction and Growth of Industrial Hemp (<i>Cannabis sativa</i>L.)

Historically, industrial hemp (Cannabis sativa L.) has been a valuable source of metabolites and compounds, such as cannabidiols. There is a need for large amounts of plant tissue to be grown under controlled environments, and plant tissue culture is one unique way to yield this tissue. The purposes of this study were to determine: 1) the optimal concentrations (μM)/ratios of auxin:cytokinin in media and;2) the optimal mineral salts formulation for callus induction and callus growth in select hemp cultivars. To find the optimal concentration/ratios, 16 different combinations of auxin:cytokinin and three different mineral salts formulations were evaluated. The three mineral salts formulations tested were MS salts, MB5D1K and an MTSU formulation. The top performing hormone formulations were determined to be equal concentrations (1:1, 2:2, 3:3 μM) of auxin and cytokinin. The top performing media hormone formulations for callus induction were determined to be 2:1, 2:2, 2:3, and 3:2 μM (auxin:cytokinin). The optimal mineral salts formulation was determined to be MD5D1K. Therefore, the overall optimal media formulation for hemp callus production would be MB5D1K salts with the concentration/ratio of 2:2 μM (auxin:cytokinin).

水稻早衰突变体psl4(t)的表型鉴定及基因定位

水稻叶片早衰严重影响水稻的产量与品质,挖掘水稻叶片早衰相关基因并解析其分子机制,对提高水稻产量具有重要意义.从涪恢9802和LR杂交后代中筛选到叶早衰突变体psl4(t),该突变体6叶期前叶片呈正常绿色,从7叶期至剑叶(倒1叶)期每张叶片均是从叶尖至叶中部逐渐衰老,叶片的叶绿体发育受阻其体积变小、光合色素质量分数减少,叶片提前衰老.农艺性状分析结果表明:与野生型相比,突变体psl4(t)的穗长、有效分蘖数和籽粒宽变化无统计学意义,而株高、每穗粒数、结实率及千粒质量显著降低.遗传分析结果发现:突变体psl4(t)的早衰性状受单隐性核基因控制,利用分子标记将目标基因定位于第4染色体长臂端两个SSR标记(RM17004和RM17006)之间38.5 kb的范围内.研究为psl4(t)基因的克隆及功能解析、早衰分子机制研究奠定基础.

<i>Cannabis sativa</i>L. Response to Narrow Bandwidth UV and the Combination of Blue and Red Light during the Final Stages of Flowering on Leaf Level Gas-Exchange Parameters, Secondary Metabolite Production, and Yield

Many modifications to growing environments and specific stress induction techniques exist which are supported by preliminary research to increase yield and/or concentration of secondary metabolites in Cannabis sativa L. One such technique is the modification of the spectral composition of light late in flowering-stage development. In this study, we evaluated the hypothesis that the addition of narrow bandwidth UV-A light and the combination of narrow bandwidth blue and red light, during the final two weeks of the flowering cycle affect the concentration of cannabinoids and terpenes, with no difference in harvest dry weight. Three treatments were used in this experimental design, 1) control-full spectrum illumination, 2) UV-A (~390 nm) at ~66 PPFD in addition to control lighting, and 3) blue (~450 nm) and red (~660 nm) at ~158 PPFD in addition to control lighting. Light treatments were initiated during the final two weeks of flowering to assess secondary metabolite concentration (cannabinoids and terpenes) and yield of three cannabis cultivars (Type I (Larry OG), Type II (Pootie Tang), and Type III (Super White)). With two of the three cultivars used in this study responding with enhanced production of THC our results support the potential beneficial effects of attributed light-mediated treatments on cannabinoid production. On the other hand, our results show contrasting trends for terpene production and yield of dry weight flower.

Study on the Anti-inflammatory and Repairing Effects of Cannabis Sativa Leaf Extract by Epikutis Model Substitution

To explore the anti-inflammatory effect of Cannabis sativa leaf extracts and their repair to damaged skin,a 3D epidermal model(Epikutis®)was used.During the experiment,after epithelial cell surface was pretreated with SLS for 24 h,the anti-inflammatory effect and repair effect of Cannabis sativa leaf extract were observed by testing tissue viability and the representative pro-inflammatory factors such as IL-1α,TNF-αand IL-8.The tissue viability was detected by the MTT test.The results showed that the Cannabis sativa leaf extract effectively inhibited the secretion of inflammatory factors IL-1α,TNF-αand IL-8,and the tissue viability of the skin model was significantly improved after using the sample.The Cannabis sativa leaf extract has a repairing effect on skin barrier damage by inhibiting skin inflammatory reaction.

亚热带地区水稻(Oryza sativa L.)气孔臭氧通量和产量的响应关系

基于开放式臭氧浓度升高O_3-FACE(Free-Air Concentration Elevation of O_3)实验平台,利用前期水稻O_3-FACE试验的基础数据,通过建立水稻产量与不同评价指标(累积气孔O_3吸收通量PODY和O_3浓度指标AOTX)的响应关系,比较了水稻产量损失与各评价指标的相关性差异,通过对暴露剂量、吸收通量相关参数取值与产量损失的观察和分析结果的比较,找出更为合理的农作物臭氧风险评估阈值。结果表明:随着通量阈值Y[0~11 nmol O_3·m^(-2)PLA·s^(-1)(PLA:projected leaf area,投影叶面积)]和暴露浓度阈值X(0~50 n L·L^(-1))的增加,回归分析R^2值逐渐增加,当Y为11 nmol O_3m^(-2)PLA·s^(-1)和X为50 n L·L^(-1)时,气孔臭氧吸收通量POD11和累积暴露剂量AOT50与水稻相对产量的相关性最大,当通量阈值Y为8~13 nmol O_3·m^(-2)PLA·s^(-1)和暴露阈值X为46~58 n L·L^(-1)时,可获得较高的R^2值取值范围,分别为0.70~0.75和0.70~0.745。参考文献发现,目前地表臭氧污染可能引起的水稻产量损失范围为5%~8%,对照圈中POD9~10和AOT40~45产量损失的预测值亦在这区间,但前者R^2值(0.73~0.74)明显高于后者R^2值(0.64~0.69),表明基于气孔臭氧通量的评价指标能更好地反映水稻产量的变化。通过进一步分析发现,当通量阈值Y为9 nmol O_3·m^(-2)PLA·s^(-1)时,能更准确地评估水稻产量损失,且其R^2值(0.73)高于通量指标POD6(0.57)。以上研究结果表明,通量指标POD9更适合评估亚热带地区O_3污染对水稻作物的影响。

水稻(Oryza sativa L·)捕光叶绿素a/b结合蛋白基因全长cDNA的克隆和特性分析

根据捕光叶绿素a／b结合蛋白基因（cab）家族中的保守序列设计PCR引物，扩增出的约310bpcDNA小片段为分子杂交探针，对构建的水稻cDNA文库进行杂交筛选，并结合PCR分析确定阳性克隆中cDNA片段的大小。通过对插入的cDNA片段最长的阳性克隆进行测序分析验证，克隆了水稻中1个cab基因全长cDNA，命名为cab-n8（GenBank登记号：AY445626）。cab-n8长为1128bp，从第55bp开始至789bp含有1个开放阅读框和1个终止密码子，编码244个氨基酸（GenBank登记号为AAR19267．1）；在3’端含有330bp的非编码区和Poly（A）18；在5’端有45bp的非编码区，在转录起始位点附近有TCA序列。通过序列分析，cab-n8编码的蛋白质在第54—216位包括典型的捕光叶绿素a／b结合蛋白功能域（chlorophll a／b binding domain）；在第141—158位含有无名指结构功能域（ring finger structure domain），该位点可能与捕光叶绿素结合蛋白与叶绿素a／b的结合有关；在第194—231位含有甘油醛-3-磷酸脱氢酶C端功能域（C-terminal domain of glyceraldehyde-3-phosphate dehydrogenase-like）。cab-n8编码的蛋白质预测的等电点和分子量分别为6．52和26955．80 Da。通过比较分析，cab-n8DNA序列（AY445626）和编码的氨基酸序列（AAR19267．1）与cab27DNA序列（AF094775．1）和编码的氨基酸序列（AAC67557．1）相似性最高，均为97％，显示cab家族基因在进化过程中是相当保守的。Northern blot分析表明，该基因在水稻叶和茎中表达没有差异，但光对其表达有明显的诱导促进作用。