重组谷糠源过氧化物酶的克隆、表达及逆转结肠癌化疗耐药活性

本课题组前期首次从谷糠中获得具抗肿瘤活性的过氧化物酶FMBP。为了该蛋白质后期在体外的规模化生产,本研究以谷子cDNA为模板,利用基因工程手段构建了pMal-s-FMBP融合质粒,并优化诱导条件进行原核表达与纯化,最终获得纯度大于90 %的具抗肿瘤活性的重组FMBP（Re-FMBP）。研究发现,Re-FMBP可以逆转HCT-8/5-Fu耐药细胞对5-Fu的耐药性,耐药逆转倍数达到9.6倍。通过Annexin V/碘化丙啶双染色流式细胞仪检测显示：Re-FMBP能够诱导HCT-8/5-Fu耐药细胞发生凋亡;通过罗丹明123染色,流式细胞仪检测结果显示,Re-FMBP处理后,HCT-8/5-Fu耐药细胞内的荧光强度增强。说明Re-FMBP能够增加5-Fu药物在HCT-8/5-Fu耐药细胞内的蓄积。结果揭示,Re-FMBP蛋白可通过抑制耐药细胞增殖,诱导耐药细胞凋亡,抑制耐药细胞对5-Fu药物外排功能来增加HCT-8/5-Fu耐药细胞对5-Fu药物的敏感性。因此,谷糠来源的Re-FMBP蛋白可以作为肿瘤化疗辅助药物来开发。     

Fringes of the empire: Diet and cultural change at the Roman to post‐Roman transition in NW Iberia

A growing number of paleodiet investigations over recent years have begun to reveal the stark dietary differences that existed between regions of the Roman Empire, as well as significant changes in subsistence strategies after its fall. The present study explores the dietary changes at the Roman to post‐Roman (Germanic) transition in the Northwest Iberian Peninsula, in order to improve our understanding of the changes that occurred at end of the Roman Empire in different regions across Europe and to also consider the influence of climate had on them. The carbon and nitrogen stable isotope investigation in bone collagen from A Lanzada, NW Spain (100–700 AD), which was an important commercial, coastal settlement has been presented. A human sample of 59 individuals, 6 of them subadults, is compared with 31 faunal specimens, which include a number of marine fish. Isotope data for the terrestrial fauna reveal the influence of the sea on the local isotope baseline. Analysis of the human samples indicates a mixed marine‐terrestrial diet. A shift in mean human δ¹³C values from ?16.7‰ to ?14.3‰ provides clear evidence for a significant change in diet in the post‐Roman period, probably through the intensification of both marine resources exploitation and C₄‐plant consumption (presumably millet). A deterioration of paleoenvironmental conditions, together with a poor socioeconomic situation and the arrival of new people, the Sueves, who brought a new political and socioeconomic system have been discussed as the main causes for the dietary modification in post‐Roman times.     

Ability of <Emphasis Type="Italic">Emericella rugulosa</Emphasis> to mobilize unavailable P compounds during Pearl millet [<Emphasis Type="Italic">Pennisetum glaucum</Emphasis> (L.) R. Br.] crop under arid condition

Phosphate solubilizing microorganisms are ubiquitous in soils and could play an important role in supplying P to plants where plant unavailable P content in soil was more. A phosphatase and phytase producing fungus Emericella rugulosa was isolated and tested under field condition (Pearl millet as a test crop) in a loamy sand soil. In the experimental soil 68% organic phosphorous was present as phytin; less than 1% of phosphorous was present in a plant available form. The maximum effect of inoculation on different enzyme activities (acid phosphatase, alkaline phosphatase, phytase, and dehydrogenase) was observed between 5 and 8 weeks of plant age. The depletion of organic P was much higher than mineral and phytin P. The microbial contribution was significantly higher than the plant contribution to the hydrolysis of the different P fractions. A significant improvement in plant biomass, root length, seed and straw yield and P concentration of root and shoot resulted from inoculation. The results suggest that Emericella rugulosa produces phosphatases and phytase, which mobilize P and enhance the production of pearl millet.     

Transgenic and herbicide resistant pearl millet (Pennisetum glaucum L.) R.Br. via microprojectile bombardment of scutellar tissue

Four different pearl millet breeding lines were transformed and led to the regeneration of fertile transgenic plants. Scutellar tissue was bombarded with two plasmids containing the bar selectable marker and the -glucuronidase reporter gene (gus or uidA) under control of the constitutive CaMV 35S promoter or the maize Ubiquitin1 promoter (the CaMV 35S is not a maize promoter). For the delivery of the DNA-coated microprojectiles, either the particle gun PDS 1000/He or the particle inflow gun was used. The calli and regenerants were selected for their resistance to the herbicide Basta (glufosinate ammonium) mediated by the bar gene. Putative transformants were screened for enzyme activity by painting selected leaves or spraying whole plants with an aqueous solution of the herbicide Basta and by the histochemical GUS assay using cut leaf segments. PCR and Southern blot analysis of genomic DNA indicated the presence of introduced foreign genes in the genomic DNA of the transformants. Five regenerated plants represent independent transformation events and have been grown to maturity and set seed. The integration of the bar selectable and the gus reporter gene was confirmed by genomic Southern blot analysis in all five plants. All five plants had multiple integrations of both marker genes. To date, the T₁ progeny of three out of four lines generated by the PDS particle gun shows co-segregating marker genes, indicating an integration of the bar and the gus gene at the same locus in the genome.     

Tuareg, a novel miniature-inverted repeat family of pearl millet (Pennisetum glaucum) related to the PIF superfamily of maize

Miniature-inverted repeat transposable elements (MITEs) are abundantly repeated in plant genomes and are especially found in genic regions where they could contribute regulatory elements for gene expression. We describe with molecular and cytological tools the first MITE family reported in pearl millet: Tuareg. It was initially detected in the pearl millet ortholog of Teosinte-branched1, an important developmental gene involved in the domestication of maize. The Tuareg family was amplified recently in the pearl millet genome and elements were found more abundant in wild than in domesticated plants. We found that they shared similarity in their terminal repeats with the previously described mPIF MITEs and that they are also present in other Pennisetum species, in maize and more distantly related grasses. The Tuareg family may be part of MITEs activated by PIF-like transposases and it could have been mobile since pearl millet domestication. Electronic supplementary material Electronic supplementary material is available for this article at and accessible for authorised users. O. Robin contributed the FISH and fiber-FISH hybridizations.     

Hypersensitive response, cell death and histochemical localisation of hydrogen peroxide in host and non-host seedlings infected with the downy mildew pathogen Sclerospora graminicola

Hypersensitive response, cell death and release of hydrogen peroxide as measures of host and non‐host defense mechanisms upon inoculation with the downy mildew pathogen Sclerospora graminicola were studied histochemically at the light microscopy level. The materials consisted of coleoptile tissues of the highly susceptible (cv. HB3), highly resistant (cv. IP18293) and induced resistant pearl millet host seedlings and non‐host sorghum (cv. SGMN10/8) and cotyledon of french bean (cv. S9). Resistance up to 80% protection against the downy mildew pathogen was induced in the highly susceptible HB3 cultivar of pearl millet by treating the seeds with 2% aqueous leaf extract of Datura metel for 3 h. Time course study with the pathogen inoculated highly resistant pearl millet cultivar revealed the appearance of hypersensitive response in 20% of seedlings as necrotic spots as early as 2 h after inoculation. In contrast, a similar reaction was observed in the highly susceptible pearl millet cultivar only 8 h after inoculation with the pathogen. In induced resistant seedlings, appearance of hypersensitive response was recorded 4 h after inoculation. Delayed hypersensitive response was observed in both the non‐host species at 10 h after inoculation. Hypersensitive response in the seedlings of the highly resistant pearl millet cultivar 24 h after inoculation showed 100% hypersensitive response, which was not observed in susceptible and non‐host species, although the induced resistant seedlings showed 90% hypersensitive response after that period of time. Cell death in the tissues of the test seedlings was also observed to change with time. Statistical analysis revealed that the tissues of highly resistant pearl millet seedlings required 2.9 h to attain 50% cell death. Tissues of induced resistant and highly susceptible pearl millet seedlings required 4.65 and 6.50 h respectively. In non‐hosts, 50% cell death was not recorded. Quantification of hydrogen peroxide in the tissue periplasmic spaces of the test seedlings revealed 2.94 h as the time required for 50% hydrogen peroxide accumulation in the tissues of highly resistant pearl millet seedlings. Tissues of induced resistant and highly susceptible pearl millet seedlings needed 3.76 and 5.5 h respectively. Fifty percent hydrogen peroxide localisation in non‐hosts could not be recorded. These results suggested the involvement of hydrogen peroxide, cell death and hypersensitive response in pearl millet host defense against S. graminicola.     

PAC对谷子花后土壤氮素供应和叶片抗氧化特性的影响

全基施肥方式会造成作物全生育期内营养供应失衡, 导致生育后期缺氮早衰。为探究聚天门冬氨酸和壳聚糖复配剂(PAC)保障谷子(Setaria italica)花后氮素供应和调控叶片抗氧化特性的机制, 建立全基施肥背景下东北春谷防衰增产的生产技术, 于2020-2021年在中国农业科学院作物科学研究所公主岭试验站开展大田试验, 以谷子品种张杂谷13号和华优谷9号为材料, 设置常规氮素(CN)和PAC配合氮素(PN) 6个氮素水平(0、75、112.5、150、225和337.5 kg·hm^-2)播种前进行全基施肥处理。结果表明, 与常规氮肥处理相比, 相同施氮量下, PAC处理后, 两品种谷子花期和灌浆中期0-20 cm和20-40 cm土层土壤硝态氮和铵态氮含量升高, 花后叶面积显著增大, 叶面积降幅减小; 花后0-40天旗叶超氧化物歧化酶、过氧化物酶及过氧化氢酶活性升高, 丙二醛含量降低。因此, PAC有效保障了谷子生育中、后期土壤氮素的供应, 提高了叶片抗氧化能力, 延缓了叶片衰老进程, 进而提高产量。2020年和2021年Z13的增产幅度分别为11.24%-21.55%和8.65%- 14.22%, H9的增产幅度分别为5.53%-15.75%和10.43%-16.17%。上述调控效应在低氮和中氮水平(75、112.5和150 kg·hm^-2)下更为显著。综上, PAC配合氮肥全基施可作为一项防衰增产的栽培技术应用于我国东北春谷区。     

山西糜子核心种质分子身份证构建

为快速鉴定糜子(Panicum miliaceum)资源, 建立分子标记检测平台, 以272份山西糜子核心种质为研究材料, 利用85对简单重复序列(SSR)引物, 应用ID Analysis 4.0软件, 构建DNA分子身份证。结果表明, 对85对SSR引物进行筛选, 发现20对引物组合(RYW67、RYW53、RYW37、RYW65、RYW62、RYW77、RYW5、RYW49、RYW84、RYW19、RYW11、RYW40、RYW54、RYW28、RYW31、RYW7、RYW16、RYW8、RYW9和RYW18)可区分272份材料。共检测到等位变异(Na) 60个, 平均每个位点检出3个; Shannon多样性指数(I)为0.957 8 (RYW16)-1.096 7 (RYW5), 平均值为1.055 2; 多态性信息含量(PIC)为0.604 4 (RYW77)-0.753 0 (RYW37), 平均值为0.692 1。利用20对引物构建山西糜子核心种质的字符串、条形码和二维码DNA分子身份证, 可为种质身份标识和溯源提供实践路径。