Characterization of a Novel Chlorophyll-Deficient Mutant Mt6172 in Wheat

Identification of new chlorophyll-deficient mutants will provide materials for studying signaling components and pathways between plastid and nucleus. A novel chlorophyll-deficient mutant, named Mt6172, was obtained by spaceflight environment induction. Genetic analysis showed that its inheritance was controlled by nuclear and cytoplamic genes. Leaf color of its self-fertilized progenies was albino, narrow-white striped, or green. Only a few cells with abnormal chloroplasts were observed in albino plants and white section of narrow-white striped plants. These chloroplasts had obvious flaws in inner structure, and granum lamellae was extremely disordered. The narrow-white striped plants were characterized with green- and-narrow-white striped leaves, and the width of stripes between different plants was even, their plant height, number of productive tillers, and 1 000-grain weight were lower than those of the wild type. The narrow-white striped plants and the wild type had significant difference in the value of potential activity of photosystem II at all tested stages. At elongation stage, which was impacted the most seriously, effective quantum yield significantly decreased, whereas the energy for photoprotection and photodamage significantly increased. changes of electron transport rate, photochemical dissipation, Under different photosynthetic active radiation conditions, and effective quantum yield were different, electron transport rate was more impacted than other parameters. Therefore, the leaf morphology and inheritance of mutant Mt6172 was different from the other reported mutants in wheat, and it was a novel mutant of chloroDhvll deficiency.

Genetic Analysis and Molecular Mapping of a Novel Chlorophyll-Deficit Mutant Gene in Rice

A rice etiolation mutant 824ys featured with chlorophyll deficiency was identified from a normal green rice variety 824B. It showed whole green-yellow plant from the seedling stage, reduced number of tillers and longer growth duration. The contents of chlorophyll, chlorophyll a, chlorophyll b and net photosynthetic rate in leaves of the mutant obviously decreased, as well as the number of spikelets per panicle, seed setting rate and 1000-grain weight compared with its wild-type parent. Genetic analyses on F1 and F2 generations of 824ys crossed with three normal green varieties showed that the chlorophyll-deficit mutant character was controlled by a pair of recessive nuclear gene. Genetic mapping of the mutant gene was conducted by using microsatellite markers and F2 mapping population of 495R/824ys, and the mutant gene of 824ys was mapped on the short arm of rice chromosome 3. The genetic distances from the target gene to the markers RM218, RM282 and RM6959 were 25.6 cM, 5.2 cM and 21.8 cM, respectively. It was considered to be a new chlorophyll-deficit mutant gene and tentatively named as chill（t）.

Photosynthesis and Dry Matter Accumulation in Different Chlorophyll-Deficient Rice Lines

Three different chlorophyll-deficient rice isogenic lines chl,fgl and pgl,and their recurrent parent zhefu802 （zf802） were used to study effects of leaf color on photosynthesis,dry matter accumulation,yield,and quality in early season indica rice.Analysis showed that the chlorophyll （Chl.） a/b ratio of isogenic lines chl-8,pgl and fgl was 5.35,10.00 and 15.46,respectively,among them,line fgl had higher leaf area index （LAI）,higher net photosynthetic rate and higher grain-filling rate than its recurrent parent zf802 at the later period of grain filling stage;while LAI,net photosynthetic rate and dry matter accumulated in lines chl-8 and pgl were lower than in zf802.Differences were found in the grain yield and quality among chlorophyll deficient isogenic lines,lines fgl,chl-8 and zf802 had similar grain yield,which was significantly higher than that of pgl;the highest milling quality was observed in isogenic line fgl,with relatively high protein content.This study showed that isogenic line fgl would become a unique material for the development of high yield rice with high grain quality because of its slow aging process and relative steady grain-filling rate.

A novel transcription factor FnMYB4 regulates pigments metabolism of yellow leaf mutants in Fragaria nilgerrensis

The strawberry species Fragaria nilgerrensis Schlechtendal ex J.Gay,renowned for its distinctive white,fragrant peach-like fruits and strong disease resistance,is an exceptional research material.In a previous study,an ethyl methane sulfonate(EMS)mutant library was established for this species,resulting in various yellow leaf mutants.Leaf yellowing materials are not only the ideal materials for basic studies on photosynthesis mechanism,chloroplast development,and molecular regulation of various pigments,but also have important utilization value in ornamental plants breeding.The present study focused on four distinct yellow leaf mutants:mottled yellow leaf(MO),yellow green leaf(YG),light green leaf(LG),and buddha light leaf(BU).The results revealed that the flavonoid content and carotenoid-to-chlorophyll ratio exhibited a significant increase among these mutants,while experiencing a significant decrease in chlorophyll and carotenoid contents compared to the wild type(WT).To clarify the regulatory mechanisms and network relationships underlying these mutants,the RNA-seq and weighted gene coexpression network(WGCNA)analyses were employed.The results showed flavonoid metabolism pathway was enriched both in MO and YG mutants,while the chlorophyll biosynthesis pathway and carotenoid degradation pathway were only enriched in MO and YG mutants,respectively.Subsequently,key structural genes and transcription factors were identified on metabolic pathways of three pigments through correlation analyses and quantitative experiments.Furthermore,a R2R3-MYB transcription factor,FnMYB4,was confirmed to be positively correlated with flavonoid synthesis through transient overexpression,virus-induced gene silencing(VIGS),and RNA interference(RNAi),accompanying by reoccurrence and attenuation of mutant phenotype.Finally,dual-luciferase(LUC)and yeast one-hybrid assays confirmed the binding of FnMYB4 to the FnFLS and FnF3H promoters,indicating that FnMYB4 positively regulates flavonoid synthesis.In addition,correlation analyses suggested that FnMYB4 also might be involved in chlorophyll and carotenoid metabolisms.These findings demonstrated the pivotal regulatory role of FnMYB4 in strawberry leaf coloration.

Detection of ARV-Resistant Mutants in HIV-1-Infected Individuals in a Context of Systematic Switching to an Association Based on Dolutegravir in Abidjan, Côte d’Ivoire

The emergence of antiretroviral resistance mutations represents a major threat to the achievement of national and global goals for the elimination of HIV-1 infection. The global strategy in 2019 in Cte d'Ivoire is a new national policy for the management of people living with HIV with the administration of dolutegravir (DTG)-based fixed-dose combination. The aim of our study was to evaluate HIV-1 resistance to antiretrovirals (ARVs) in infected adult subjects in Cte d’Ivoire in the context of a systematic switch to a DTG-based combination. Between February 2022 and October 2023, a cross-sectional survey with random sampling was conducted in 06 services caring for people living with HIV. A total of 139 participants were included in the study. Adults with a viral load ≥ 1000 copies/mL were tested for HIV-1 ARV resistance mutations. Molecular analyses were performed using protocol of ANRS-MIE (National Agency for Research on AIDS and emerging infectious diseases). The interpretation is performed by HIVGRAD (https://www.hiv-grade.de/cms/grade/). The frequencies of HIV-1 resistance to non-nucleotide reverse transcriptase inhibitors (NNRTIs), nucleotide reverse transcriptase inhibitors (NRTIs), integrase inhibitors (IINTs) and protease inhibitors (PIs) were 82%, 73%, 19% and 11% respectively. The main mutations observed in the different classes were K103N (45%), M184V (64%), E157Q (19%) and L10V/M46I/A71V/I54V (6%) respectively. This study reveals the emergence of resistance to DTG-based fixed-dose combinations, favored by high rates of resistance to NRTIs and NNRTIs. This finding underlines the need for enhanced viral load monitoring and HIV-1 genotyping tests to guide the choice of NRTIs for combination therapy. In addition, monitoring for mutations to second-generation NRTIs is essential, given the scale-up of DTG-based regimens currently underway in Cte d’Ivoire.

An efficient method for constructing a random insertional mutant library for forward genetics in Nannochloropsis oceanica

Insertional mutation,phenotypic evaluation,and mutated gene cloning are widely used to clone genes from scratch.Exogenous genes can be integrated into the genome during non-homologous end joining(NHEJ)of the double-strand breaks of DNA,causing insertional mutation.The random insertional mutant library constructed using this method has become a method of forward genetics for gene cloning.However,the establishment of a random insertional mutant library requires a high transformation efficiency of exogenous genes.Many microalgal species show a low transformation efficiency,making constructing random insertional mutant libraries difficult.In this study,we established a highly efficient transformation method for constructing a random insertional mutant library of Nannochloropsis oceanica,and tentatively tried to isolate its genes to prove the feasibility of the method.A gene that may control the growth rate and cell size was identified.This method will facilitate the genetic studies of N.oceanica,which should also be a reference for other microalgal species.

Roles of Mutant TP53 Gene in Cancer Development and Progression

TP53 is a tumor suppressor gene that is mutated in most cancer types and has been extensively studied in cancer research.p53 plays a critical role in regulating the expression of target genes and is involved in key processes such as apoptosis,cell cycle regulation,and genomic stability,earning it the title“guardian of the genome.”Numerous studies have demonstrated p53’s influence on and regulation of autophagy,ferroptosis,the tumor microenvironment,and cell metabolism,all of which contribute to tumor suppression.Alterations in p53,specifically mutant p53(mutp53),not only impair its tumor-suppressing functions but also enhance oncogenic characteristics.Recent data indicate that mutp53 is strongly associated with poor prognosis and advanced cancers,making it an ideal target for the development of novel cancer therapies.This review summarizes the post-translational modifications of p53,the mechanisms of mutp53 accumulation,and its gain-of-function,based on previous findings.Additionally,this review discusses its impact on metabolic homeostasis,ferroptosis,genomic instability,the tumor microenvironment,and cancer stem cells,and highlights recent advancements in mutp53 research.

Integrated metabolomics and proteomics analyses reveal the molecular mechanism underlying the yellow leaf phenotype of Camellia sinensis

The tea plant cultivar‘Zhonghuang 2'(ZH2)possesses albino-induced yellow leaves that contain low levels of catechins but high contents of amino acids.However,the molecular mechanism underlying the yellow leaf phenotype of ZH2 has not been elucidated clearly.In the current research,the yellow shoots(ZH2-Y)and naturally converted green shoots(ZH2-G)of ZH2 were studied using metabolic and proteomic profiling for a better understanding of the mechanism underlying phenotype formation.In total,107 differentially changed metabolites(DCMs)were identified from the GC-MS-based metabolomics,and 189 differentially accumulated proteins(DAPs)were identified from the tandem mass tag(TMT)-based quantitative proteomics.Subsequently,integrated analysis revealed that‘porphyrin and chlorophyll metabolism',‘carbon fixation in photosynthetic organisms',and‘phenylpropanoid biosynthesis'pathways were commonly enriched for DAPs and DCMs.We further found that the inhibition of chlorophyll biosynthesis,the deficiency of photosynthetic proteins and the imbalance of the ROS-scavenging system were the crucial reasons responsible for the chlorosis,chloroplast abnormality and photooxidative damage of ZH2 leaves.Altogether,our research combines metabolomics and proteomics approaches to uncover the molecular mechanism leading to the yellow leaf phenotype of tea plants.

搭载实践十号卫星的不同小麦品种突变体鉴定与分析

为探讨卫星搭载对小麦农艺性状、高分子量麦谷蛋白亚基(HMW-GS)和分子水平变异的影响,本研究利用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)技术、简单重复序列(SSR)标记技术对实践十号返回式科学实验卫星搭载周麦18、周麦22和汶农14小麦品种干种子的SP3代农艺性状变异和SP5代突变系的HMW-GS和SSR分子标记多态性进行鉴定与分析。结果发现,航天诱变SP3代农艺性状与其野生型存在明显差异,且不同小麦品种对太空环境的敏感性不同。航天搭载能够诱发高分子量麦谷蛋白亚基和基因组DNA发生变异,发现3个小麦品种高分子量麦谷蛋白亚基的变异频率分别为2.15%、3.66%和5.21%,其中汶农14突变体HMW-GS亚基变异频率最高。21个SSR分子标记检测结果显示,周麦18和周麦22的航天诱变突变体与其野生型差异标记数量均不超过2个,而汶农14航天诱变的部分突变体与其野生型间差异标记数量较多,且株高和穗型等性状发生了遗传分离。综上,航天诱变使小麦基因组和蛋白组发生变异,所创制的优异特色突变体可为小麦育种的遗传改良和功能基因的研究利用提供丰富的材料基础。

Antioxidase System of a Rice Mutant with Low Chlorophyll b

[Objective] The mitigative effect of antioxidase system of a rice mutant with low chlorophyll b on photooxidative damage was studied.[Method] A rice mutant with low chlorophyll b and its wild type were taken as experimental materials to comparatively research their peroxide (H2O2) contents, the activity and isozymes of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) in chloroplast.[Result] Compared with the wild type, there were many kinds of SOD, POD and CAT isozymes in leaf cells and chloroplast cell of mutant, and the activity of SOD, POD and CAT isozymes in leaf cells and chloroplast cell of mutant was also correspondingly higher. Under intense light condition, the H2O2 content of chloroplast in mutant was less than that in the wild type. [Conclusion] The higher activity of scavenging active oxygen can relieve the photooxidative damage made by excessive light energy of intense light on photosynthetic membrane, which is an important reason for higher photosystem Ⅱ (PS II) stability of this mutant.

Genetic Analysis of a Small Grain Mutant Induced by T-DNA Insertion in Rice

[Objective] The aim of this study is to understand the genetic characteristics of a grain shape mutant and its possible role in genetic improvement of grain yield in rice. [Method] On the basis of the collection of T-DNA tag lines, the progeny of homozygous plants carrying T-DNA insertion were screened for mutants with mutated phenotypes. The genetic analysis of the mutant and test for the linkage between the mutated phenotype and the T-DNA insertion were carried out to determine its genetic characteristics. [Result] In the present study, a grain shape mutant induced by T-DNA insertion in rice was identified, which showed small grain. Genetic analysis of the mutant showed that the two types of phenotype, normal and small grain in the segregating populations derived from the T-DNA heterozygotes, fit the ratio of 3∶1. Test for Basta resistance showed that all the mutants were resistant while the normal plants segregated for resistant and susceptible by the ratio of 2∶1. The results indicated that the mutant phenotype cosegregated with Bar gene. The small grain mutant caused by T-DNA insertion was confirmed by PCR amplification aiming at T-DNA. [Conclusion] The grain shape mutant is useful for isolation of the tagged gene and genetic improvement in rice.

Study on a Mutant with Low Content Chlorophyll b in a High Yielding Rice and Its Photosynthesis Properties

A high yielding rice mutant ( Oryza sativa L. cv. Zhenhui 249) with low chlorophyll b was recently discovered in the field. The mutant was mainly characterized by the decrease of the content of extrinsic antennae complex. This variation was shown in the stage when the leaves were expanding. When the leaves are at the final developmental stage, the content would approach to that of the wild type. It was discovered that only moderate amount of chlorophyll b decreased in this mutant. The photosynthetic apparatus of the mutant was rather stable in the whole life span of the leaf. The extrinsic antennae complex of the mutant might make efficient use of light and meanwhile reduce the production of O -· 2.

Physiological Character and Gene Mapping in a New Green-revertible Albino Mutant in Rice

A green-revertible albino mutant-Qiufeng M was found from the japonica rice （Oryza sativa L. ssp. japonica） Qiufeng in the field. The first three leaves of the mutant were albino with some green. The leaf color became pale green since the fourth leaf and the glume had the same phenomenon as the first three leaves. The measuring data of the pigment content confirmed the visually observed results. It truly had a remarkable changing process in the leaf color in Qiufeng M. Comparison of the main agronomic characters between Qiufeng and Qiufeng M indicated that the neck length and grain weight showed significant difference at the 1% level, and other characters were not different. Genetic analysis showed that the green-revertible albino trait was controlled by a single recessive nucleic gene. Using 209 recessive mutant individuals in the F2 population derived from the cross Pei＇ai 64S × Qiufeng M, a gene, tentatively named gra（t）, was located between the SSR markers of RM475 and RM2-22 on the long arm of chromosome 2. The genetic distance were 17.3 cM and 2.9 cM respectively.

Genetic Analysis of a Biomass Mutant in Oryza sativa

[ Objective ] The study aimed to reveal the genetic model of a biomass mutant in Oryza sativa. [ Method ] In the process of screening and identification of Bar-transgenic rice, a biomass mutant was found in 10 lines of T1 progenies. The mutant was investigated for genetic analysis and agronomic traits by herbicide spraying and PCR amplification. [ Result] The segregation ratio is consistent with mendelian law（3：1）. The mutant assumed not only higher plant height, wider straw and earlier florescence, but also more tillers, bigger spikes and resultantly higher biomass. PCR detections indicated that no co-segregation was observed between mutant traits and target gene（Bar） in the T-DNA inserted, proving that the mutant is not caused by the insertion of T-DNA containing target gene （Bar）. [ Conclusion] Our study may avail to understand the cloning of mutant gene and the mechanism of the mutant gene on biomass.

Chloroplast Composition and Structural Differences in a Chlorophyll-reduced Mutant of Oilseed Rape Seedlings

对黄化突变体Cr352 9和野生型油菜 (BrassicanapusL .) 352 9叶绿体的超微结构和组成进行了比较。与野生型相比 ,突变体Cr352 9叶片具有较少的类囊体、较少的垛叠膜区和较少的叶绿素含量。突变体的Chla/Chlb比值较高 ,是野生型的 2倍。电泳结果表明 ,突变体类囊体膜中LHCⅡ和其三聚体LHCⅡ 的含量减少。SDS_PAGE分析显示 ,LHCⅡ的脱辅基蛋白在突变体类囊体膜中明显减少。免疫印迹进一步表明 ,所有LHCⅡ组分的含量仅为野生油菜的类囊体膜的 1 / 3。突变体Cr352 9的天线系统比野生型 352 9的小。

Screening of Saccharomyces Strains Highly Producing Glutathione and Breeding of Its Ethionine-resistant Mutants

[Objective] The aim of this study was to screen Saccharomyces for glutathione over-production. [Method] Ethionine-resistant mutants were obtained through UV mutagenesis and rational screening. [Result] A high GSH-producing strain HSJB1 was isolated from soil, and the biomass for this strain by flask shaking fermentation was 3.87 g/L while the GSH yield was 91.87 mg/L. According to the morphological, physiological and biochemical characteristics of cells, this strain was primarily identified as Saccharomyces cerevisiae. An ethionine-resistant mutant YBS77 was obtained through UV mutagenesis of the original strain HSJB1, and the biomass for this strain by flask shaking fermentation was 7.60 g dry cell weight/L while the GSH yield was 211.96 mg/L. [Conclusion] The biomass of the mutant obtained by breeding is increased by 96.38% than that of the original strain, and the GSH yield of the mutant obtained by breeding is increased by 130.72% than that from the original strain, which indicates that the breeding method is feasible.

Construction of Mutant Population and Analysis of Dwarf Mutants in "6421"(Capsicum annuum L.)through EMS Mutagenesis

[Objective] The aim was to improve the genetic property of peppers, the mutant population of Capsicum annuum L cultivar ＂6421＂ was constructed. [Method] The seeds of ＂6421＂ were treated with 0.2% to 1.2% ethyl methane sulfonate to identified LD50, and then 10 000 LD^o of treated seeds were sowed to construct mutant population. The agronomic characters and genetic regularity of dwarf mutants in M4 generation were analyzed. [Result] Our results showed that GR and SSR were 45.2% and 40.2% respectively at 1.0% EMS, close to LD50, with GI （17.6） and seed Ⅵ （19.7） being half of that of control; 562 M4 mutants were identified in 2015, and the mutation could be characterized according 11 major categories and 32 subcategories; Simultaneously, we found that plant height, plant width, diameter of mainstem, length of main-stem, the number of main-stem nodes and branch of lines E29, E58, E142 and E312 were all significantly different from that of the control. The mutation of lines E29, E58 and E312 was all controlled by a single recessive gene. [Conclusionl The study first created a pepper mutant population, which provides not only the germplasm resources for further breeding but also direct and effective materials for genomic study of the pepper.

Physiological Analysis of Two Arabidopsis thaliana Mutants in Response to CO2

[Objective] The 15urpose was to seek for the different phenotypes between wild type and Arabidopsis Mutants in response to CO2. [Method] The epidermis bioassays and seed germination test were carried out to analyze the physiological characteristics of two Arabidopsis mu- tants and their wild type. [Result] There existed distinct differences in stomata apertures, water loss and leaf temperature compared with wild type except for stomata density. In addition, seed germination test on the medium indicated that cdfl was insensitive to ABA, mannitol and NaCI, but cdsl performed contrary to cdil. [ Conclusion] There are some different physiological characteristics between wild type and mutants.

Morphological,Anatomical and Genetic Analysis for a Rice Mutant with Abnormal Hull

A mutant with abnormal hull was first discovered from a twin-seedling strain W2555 in rice （Oryza sativa L.）. The mutant had sparse branches and decreased number of florets from the base to the peak. Frequently, the florets at the top of the panicle did not develop completely. The underdeveloped florets often showed slender and white in their life cycle. Genetic analysis indicated that the mutant traits were controlled by a single recessive gene （temporarily designated as ah）. ah gene controlled the development of inflorescence meristem and the flower organ. The florets of mutant showed degenerated lemma and palea. Stamens and lodicules were homeoticly transformed into pistils and palea/lemma-like structures, respectively. It seemed that ah mutant phenotypes of the homeotic conversions in lodicules and stamens were very similar to that of the B loss-of-function spwl gene reported previously in rice.

Grain Filling Characteristics of Giant Embryo Rice Mutant

[Objective] The aim was to explore the relevance between decreased 1 000-grain weight and grain-filling characteristics of rice giant embryo mutant.[Method] Richards equation was used to describe grain filling processes of giant embryo mutant MH-ge1 and its corresponding wild type so as to analyze the parameters of grain-filling characteristics of materials.[Result] The initial filling power of mutant MH-ge1 was higher than the wild type MH86,however,the maximum filling rate,the mean filling rate and the final increment of mutant MH-ge1 was lower than those of MH86.[Conclusion] The grain-filling characteristics of mutant MH-ge1 were poorer than the wild type MH86,thus resulting in the decrease of the grain weight of the giant embryo rice.