Less Chlorophyll Does not Necessarily Restrain Light Capture Ability and Photosynthesis in a Chlorophyll‐Deficient Rice Mutant

A chlorophyll‐deficient rice mutant, yellow‐green leaf 1 (ygl1), with a photosynthetic rate similar to that of wild type (Oryza sativa L., cv. Zhenhui 249 China) is presented in this study. Forty percent of the chlorophyll in the mutant captured 70 % of the light in photosystem II (PSII) compared with wild type. The 30 % decreased light in the mutant was compensated for by a relatively higher quantum yield of PSII, which conferred a total electron transport rate (J_T) equal to that in wild type. More photons were absorbed by wild type and exhausted through thermal dissipation. Gene expression analysis of the thylakoid membrane showed that the chlorophyll deficiency in the mutant did not impair the electron transport chain. To determine why the photosynthetic rate was similar between the chlorophyll‐deficient mutant and wild type, the leaf nitrogen (N) and Rubisco contents, stomatal and mesophyll CO₂ conductance, and chloroplast development were investigated. Our results indicate that the chlorophyll deficiency in the mutant had no negative impact on the chloroplast development in terms of size and grana stacking; moreover, the chloroplastic CO₂ concentration and Rubisco content were comparable to those in wild type. We conclude that the light‐capturing ability of normal rice plants is not fully utilized and that absorbed light in the chlorophyll‐deficient mutant is more efficient in participating photosynthesis.     

Characterization and mapping of novel chlorophyll deficient mutant genes in durum wheat

The yellow-green leaf mutant has a non-lethal chlorophyll-deficient mutation that can be exploited in photosynthesis and plant development research. A novel yellow-green mutant derived from Triticum durum var. Cappelli displays a yellow-green leaf color from the seedling stage to the mature stage. Examination of the mutant chloroplasts with transmission electron microscopy revealed that the shape of chloroplast changed, grana stacks in the stroma were highly variable in size and disorganized. The pigment content, including chlorophyll a, chlorophyll b, total chlorophyll and carotene, was decreased in the mutant. In contrast, the chla/chlb ratio of the mutants was increased in comparison with the normal green leaves. We also found a reduction in the photosynthetic rate, fluorescence kinetic parameters and yield-related agronomic traits of the mutant. A genetic analysis revealed that two nuclear recessive genes controlled the expression of this trait. The genes were designated ygld1 and ygld2. Two molecular markers co-segregated with these genes. ygld 1 co-segregated with the SSR marker wmc110 on chromosome 5AL and ygld 2 co-segregated with the SSR marker wmc28 on chromosome 5BL. These results will contribute to the gene cloning and the understanding of the mechanisms underlying chlorophyll metabolism and chloroplast development in wheat.     

Identification of DNA markers linked to an induced mutated gene conferring resistance to powdery mildew in pea (<Emphasis Type="Italic">Pisum sativum</Emphasis> L.)

We have recently induced two powdery mildew (Erysiphe pisi Syd) resistant mutants in Pisum sativum L. via ethylnitrosourea (ENU) mutagenesis. Both mutations (er1mut1 and er1mut2) affected the same locus er1 that determines most of the identified natural sources of powdery mildew resistance (PMR) in this crop. The mutated gene er1mut2 was mapped to a linkage group of 16 DNA markers combining three main strategies: near isogenic lines (NILs) analysis, bulked segregant analysis and genetic mapping of randomly identified polymorphic markers, together with three DNA-markers techniques: ISSR, RAPDs and AFLPs. Markers located closer to the PMR locus, OPO06_1100y (0.5 cM), OPT06₄₈₀ (3.3 cM) and AGG/CAA₁₂₅ (5.5 cM), were cloned and converted into SCAR markers. Markers AH1R₈₅₀ and AHR_920y were found to be allelic and converted into the co-dominant marker ScAH1 (16.3 cM). Two previously known DNA markers, ScOPE16₁₆₀₀ and A5_420y, were mapped at 9.6 and 23.0 cM from the PMR locus, respectively. The novel markers identified in this study are currently being transferred to a new F2 mapping population derived from a cross between the induced PMR mutant line F(er1mut2) and a more genetically distant susceptible line of Pisum sativum var. arvense.     

The development of a co-dominant marker for the β-conglycinin α′ subunit in soybeans

The development of soybean varieties that lack the β-conglycinin α′ subunit is an attractive goal because the β-conglycinin α′ subunit exerts a negative influence on nutrition and tofu gelation, and is also a major allergen. We sought to develop a co-dominant DNA marker for the β-conglycinin α′ subunit (Cgy-1) gene for use in marker-assisted selection (MAS). We identified a deleted sequence responsible for the null allele of Cgy-1 in a soybean variety lacking the β-conglycinin α′ subunit known as ‘Keburi’. The deletion spanned 12,998 bp and included Cgy-1 and its incomplete tandem duplicate on chromosome 10. Based on the Cgy-1 sequence from both the Williams 82 soybean reference sequence and the Keburi variety, a set of three allele-specific primers was designed for multiplex PCR assay. These primers enabled allelic discrimination by the sizes of PCR products. This amplified two distinct DNA fragments of 913 and 649 bp in Williams 82 and Keburi, respectively. The practicality of the developed co-dominant marker for Cgy-1 was also confirmed by amplification in five other soybean varieties including three wild types and two mutants. The heterozygosity of the F₁ plants at the Cgy-1 locus was ascertained using our novel co-dominant marker. This PCR-based co-dominant marker is capable of detecting the presence or absence of β-conglycinin α′ subunit for soybean marker assisted breeding system.     

水稻叶片灰白转黄突变体pyr1的鉴定与基因定位

鉴定和克隆叶色突变基因对于深入了解叶绿素合成、降解途径的关系以及植物的光合作用有着重要的作用。从EMS诱变恢复系缙恢10号后代中鉴定出1个灰白转黄突变体pyr1,该突变体在苗期部分死亡,整张叶片呈现灰白色,在不同的生育时期叶片呈现不同的颜色,直到孕穗期叶片上部和叶缘表现黄色。苗期到抽穗期突变体叶绿素含量比野生型显著或极显著降低。透射电镜观察表明,突变体与野生型细胞结构无明显差异,但叶绿体发育异常,内部大量降解,基质片层退化。遗传分析表明该性状受1对隐性基因控制,利用326株F₂隐性定位群体将PYR1基因定位在第1染色体长臂上,位于标记RM11722和Ind1之间,物理距离约92 kb,本研究为PYR1基因的图位克隆奠定了基础。     

Fine mapping and photosynthetic characteristics of the lower chlorophyll b 1 mutant in rice (Oryza sativa L.)

Chlorophylls absorb and transfer light energy to the photosynthetic system. Consequently, chlorophyll content is strongly related to crop biomass and yield. We isolated a rice spontaneous mutant, lower chlorophyll b 1 (lcb1), from a recombinant inbred line population. Under normal growth conditions, lcb1 plants produced yellow leaves with decreased total chlorophyll and chlorophyll b contents, but normal chlorophyll a content. Photosynthetic and fluorescence parameters differed between wild‐type and lcb1 plants. Compared with wild type, lcb1 had a higher electron transfer rate, a lower photochemical quenching coefficient and significantly reduced grain number, biomass and yield. A recessive nuclear gene controlled the mutant trait. Through map‐based cloning, we located the LCB1 gene in a 117.4‐kb region on the short arm of chromosome 3, close to the centromere, in a region containing 15 predicted candidate genes. None of these genes was directly related to chlorophylls or the chloroplast; therefore, lcb1 may be a mutation of a novel gene. These results will be useful for further research on the molecular mechanisms controlling biogenesis and chloroplast biochemical processes.     

Identification and fine mapping of a candidate gene for oil yellow leaf 2 conferring yellow leaf phenotype in maize

Leaf colour is an important agronomic trait for studying molecular mechanisms in chlorophyll biosynthesis and chloroplast development. Here, a novel mutant oil yellow leaf 2 (Oy2) with a typical yellow leaf phenotype at the seedling stage was identified from the mutant population derived from the maize inbred line RP125. Compared with wild type, Oy2 mutant displays decreased chlorophyll content, reduced photosynthetic capacity and impaired chloroplast structure, which is likely controlled by a single recessive gene. The Oy2 locus was then delimited into a 117 kb region on chromosome 5 harbouring four genes, amongst which the gene Zm00001d013013, encoding a magnesium chelatase subunit D, was identified as the only candidate gene associated with Oy2 mutant phenotype. Moreover, the expression levels of candidate gene Oy2 and genes associated with chlorophyll biosynthesis and photosynthesis were tested by RNA‐seq and qRT‐PCR, implying that the causal gene Oy2 playing a critical role in chlorophyll synthesis. Taken together, we propose that the causal gene Oy2 highly associated with the yellow leaf phenotype may be helpful in elucidating photosynthetic pigments biosynthesis and chloroplast development in maize.     

Allelic relationship between spontaneous and induced mutant genes for stem fasciation in chickpea

Stem fasciation is a morphological abnormality observed in plants where the stem is widened and leaves and flowers or pods are clustered at the apex. Several spontaneous mutants and one induced mutant for stem fasciation are found in chickpea (Cicer arietinum L.). This study was aimed at determining allelic relationship between spontaneous and induced mutant genes controlling stem fasciation and effects of stem fasciation on grain yield. Two spontaneous (ICC 2042 and ICC 5645) and one induced (JGM 2) stem fasciation mutants were crossed in all combinations, excluding reciprocals. The F₁ and F₂ plants from a cross between the two spontaneous mutants had fasciated stem. This indicated the presence of a common gene (designated fas1) for stem fasciation in the two spontaneous mutants. The F₁s of the crosses of the induced mutant JGM 2 with both spontaneous mutants had normal plants and segregated in a ratio of 9 normal : 7 fasciated plants in F₂. Thus, the gene for stem fasciation in the induced mutant JGM 2 (designated fas2) is not allelic to the common gene for stem fasciation in spontaneous mutants. The two genes in dominant condition produced normal non‐fasciated stem. The fasciated and the non‐fasciated F₂ plants did not differ significantly for number of pods per plant, number of seeds per plant, grain yield per plant and seed size, suggesting that it is possible to exploit the fasciated trait in chickpea breeding without compromising on yield.     

Reaction of cultivars of populus spp. to radiation induced virulent mutants of melampsora medusae

Summary Infection Type (IT) and Uredinial Number per Leaf Disk (ULD), induced on eleven cultivars of poplar in vitro, were employed to compare five radiation induced, mono-uredinial mutant lines of Melampsora medusae Thum. with the wild type race 5A from which they were derived. IT produced by the mutants was higher than (eight cultivars), similar to (two cultivars) and pronouncedly less than (IT from 4 to 1 in P. deltoides cv. 7–2) that of race 5A. Although the five mutants produced a uniform IT within a particular cultivar. ULD varied significantly between mutants on individual cultivars and there was a significant differential interaction of the mutant lines with cultivars. The ranking of mutant lines for aggressiveness on cultivars was not consistent while the differences among the mutants in mean ULD over all cultivars were not associated with the dosage level of irradiation from which they were isolated. The implications of the results in the interaction of leaf rust with poplar cultivars are discussed.     

Genetic variability of seed-quality traits in gamma-induced mutants of sunflower (<Emphasis Type="Italic">Helianthus annuus</Emphasis> L.) under water-stressed condition

Sunflower is one of the major annual world crops grown for edible oil and its meal is a potential source of protein for human consumption. It contains tocopherol that decreases potential risk of chronic diseases in human. The objectives of the current research are to assess the genetic variability and to identify AFLP markers and candidate genes associated with seed-quality traits under well-irrigated and water-stressed conditions in gamma-induced mutants of sunflower. Two mutant lines, M8-826-2-1 and M8-39-2-1, with significant increased level of oleic acid were identified that can be used in breeding programs for quality increase high oxidative stability and heart-healthy properties. The significant increased level of tocopherol in mutant lines, M8-862-1N1 and M8-641-2-1, is justified by observed polymorphism for tocopherol pathway-related gene; MCT. The most important marker for total tocopherol content is E33M50_16 which explains 33.9% of phenotypic variance. One of the most important candidate genes involving fatty acid biosynthesis, FAD2 (FAD2-1), is linked to oleic and linoleic acids content and explained more than 53% of phenotypic variance. Common markers associated with different seed-quality traits in well-irrigated and water-stressed conditions could be used for marker-assisted selection (MAS) in both conditions. Other markers, which are specific for one condition whereas linked to different traits or specific for a trait, could be useful for a given water treatment.     

Simple sequence repeats as useful resources to study transcribed genes of cotton

Microsatellites or Simple Sequence Repeats(SSRs) are informative molecular genetic markers in many crop species. SSRs are PCR-based, highly polymorphic, abundant, widely distributed throughout the genome and inherited in a co-dominant manner in most cases. Here we describe the presence of SSRs in cDNAs of cotton. Thirty one SSR primer pairs of 220 (∼14%) tested led to PCR amplification of discrete fragments using cotton leaf cDNA as template. Sequence analysis showed 25% of 24randomly selected cDNA clones amplified with different SSR primer pairs contained repeat motifs. We further showed that sequences from the SSR-containing cDNAs were conserved across G. barbadense and G. hirsutum, revealing the importance of the SSR markers for comparative mapping of transcribed genes. Data mining for plant SSR-ESTs from the publicly available databases identified SSRs motifs in many plant species,including cotton, in a range of 1.1 to4.8% of the submitted ESTs for a given species. This revised version was published online in August 2006 with corrections to the Cover Date.     

Individual and combined effects of plant height reducing genes in periwinkle

Inheritance of an induced bushy mutant (EMS 24-5) of periwinkle (Catharanthus roseus) and its genetic relationship with an earlier reported dwarf mutant (NEU 6-15) was studied by crossing it with its parental variety, Nirmal and the dwarf mutant. The bushy mutant was found to be under the control of a monogenic recessive gene (by), which was inherited independently of the recessive gene (dw ₁ ) involved in the dwarf mutant. The double mutant (bydw ₁ ) recombinant, which was recovered from the F₂ generation of the cross (NEU 6-15) × (EMS 24-5), its parental mutants and variety, Nirmal were used for determining the individual and combined effects of plant height reducing genes dw ₁ and by. Individually, the recessive genes by and dw _1, reduced plant height by 27–30, 64–68%, respectively, while together they reduced plant height by 73–77%. The double mutant recombinant was found to be 63–68 and 25–29% shorter than its parental mutants (EMS 24-5) and (NEU 6-15), respectively. The effects of the recessive gene by on plant height were discernable only after the age of 4 months while those of dw ₁ and combined effects of dw ₁ and by became apparent one to 2 weeks after germination. The double mutant recombinant was found to have higher content of total alkaloids in the roots than parental mutants as well as the variety, Nirmal.     

Cytological and microsatellite mapping of mutant genes for spherical grain and compact spikes in durum wheat

Two mutants for sphaerococcoid seed (MA 16219) and compact spike (MA 17648) were isolated from M₃ progeny of durum wheat cultivar, Altaiskaya Niva, mutagenized with chemical mutagens. The chromosomal locations of the genes involved were determined by the use of a complete set of D-genome disomic substitutions in durum cultivar, Langdon. The gene for sphaerococcoid grain, s ¹⁶²¹⁹ , was allelic to S2, located in the centromeric region of chromosome 3B in hexaploid wheat. The gene for compact spike, C ¹⁷⁶⁴⁸ , was located on chromosome 5AL distal to the Q locus. Using microsatellite markers, C ¹⁷⁶⁴⁸ and awn inhibitor B1 were located in the F₂ of LD222 × MA17648. The gene order was Xbarc319—C ¹⁷⁶⁴⁸ —Xgwm179—Xgwm126—Xgwm291—B1.     

Pleiotropic effects of male sterility genes in hybrid tomatoes (Lycopersicon esculentum Mill.)

Summary The influence of seven recessive male-sterile alleles on early marketable yield, total marketable yield and average fruit weight in heterozygous F1 hybrid tomatoes was studied. Differences were found between nearisogenic F1 hybrid pairs, in the mutants ms-14, ms-17 and ms-18 for percentage of early marketable yield; in the mutants ms-17, ms-31 and ms-47 for average fruit weight; and in the mutants ms-14, ms-17 and ms-33 for total marketable yield. These differences may be attributed to pleiotropic effects of male-sterility alleles or genes highly linked to these alleles. The direction and the magnitude of the pleiotropic effect on a specific characteristic were found to be dependent on the general genetic background of the parental lines and the specific combining ability of the F1 hybrid. Interactions found in crosses between male-sterile and male-fertile female lines and parental lines suggest that male-sterility alleles affect the general combining ability of female lines. Such differences were demonstrated to be statistically significant for percentage of early marketable yield with mutant ms-18, for average fruit weight with mutants ms-47, and for total marketable yield with mutant ms-17 and ms-33. The absence of uniformity with respect to the pleiotropic effects and occasional deviations in the general combining ability are discussed.     

Pollen Characteristics and Genetics of Induced and Spontaneous Genetic Male-Sterile Mutants in Rice

Pollen characteristics, inheritance and allelism of 23 genetic male-sterile (ms) mutants of rice (Oryza sativa L.) were investigated. Ten of the mutants were induced by ethyl methane sulfonate, one induced by ethyleneimine, seven induced by gamma rays, one induced by streptomycin, one derived from tissue culture and three were spontaneous mutants found in the field. Four pollen abortion types were observed among these ms mutants: 3 no-pollen (NP) type, 6 complete pollen abortion (CPA) type, 13 partial pollen abortion (PPA) type, and 1 stainable pollen abortion (SPA) type. Progeny tests over two years indicated that each of the mutants was inherited as a monogenic recessive. A partial diallel cross among the 23 ms mutants indicated that mutant E2 was allelic to G4, E5 was allelic to E9, N2 was allelic to N3, and that three other mutants, E3, G6 and T1, shared the same ms locus. Gene symbols ms-46 (t) through ms-63 (t) were assigned to these mutants.