Selection of Semidwarf, Early Maturing and Blast Resistant Mutants after Mutagenic Seed Treatment in Two Locally Adapted Indian Rice Cultivars

The objective of this study was to induce semidwarf, early maturing and blast resistant mutants in two adapted Indian rice cultivars. Seeds of the rice cultivars ‘Madhu Malti’ and ‘Phul Patas 72’ were treated with gamma rays and ethyl methane sulfonate (EMS) as well as a combination of gamma rays and EMS. A total of 96 dwarf (< 75 cm) and semidwarf (< 100 cm) and 104 early flowering/maturing M₂ plants were identified among the 18,060 M₂ plants observed. Sixty-eight blast resistant M₂ plants were also identified. The true mutant status of these plants has yet to be determined in progeny tests. The induction of desired mutant types in locally adapted cultivars could prove to be superior over their introduction from non-adapted foreign materials via backcrossing in that the important agronomic characteristics of adaptation to the local environment are retained in the mutants.     

Effects of dwarfing genes on yield and yield components under irrigated and rainfed conditions in wheat (Triticum aestivum L.)

Summary Near-isogenic tall (no dwarfing gene), semidwarf (Rht1 or Rht2) and dwarf (Rht1 + Rht2 or Rht3) spring wheat lines were evaluated for yield and yield components under irrigated and rainfed conditions. Under irrigated conditions, the dwarf and the semidwarf lines exhibited a significant yield advantage over the tall lines. Under rainfed conditions, the semidwarf lines outyielded the tall as well as the dwarf lines. Percent yield reduction in response to drought stress was highest with the dwarfs and lowest with the tall lines. Dry matter production of the tall lines and that of the semidwarf lines did not differ significantly and both produced significantly more dry matter than the dwarf lines under irrigated as well as rainfed conditions. Plant height and kernel weight decreased with increasing degree of dwarfness while number of kernels per spikelet, harvest index and days to heading increased under both moisture regimes. The dwarfing genes did not have any significant influence on number of tillers/m² and spikelets per spike in either moisture regime.     

Evidence for phenotypic assortative mating for flower colour in periwinkle

Two experiments were conducted with periwinkle, Catharanthus roseus, to determine the extent of natural outcrossing. Three white-flowered, fully self-fertile, monogenic recessive mutants, namely, dwarf, wavy leaf margin and curved leaf were used, together with their parental white-flowered variety, ‘Nirmal’, and a normal pink-flowered variety, PS-1. The extent of total outcrossing ranged from 43.4 to 79.3% among mutants. Outcrossing between white × white-flowered plants ranged from 28.3 to 65.3% and was two to four times greater than that between white × pink-flowered plants in the three mutants. The extent of out-crossing between white × pink-flowered plants was similar “02.2-15.0%” in all mutants and also similar to that in the normal white-flowered variety,‘Nirmal’(00.4%), where white × white flower out-crossing could not be estimated. There were no large differences in the number of seeds per fruit, percentage fruit set and germination percentage of seeds obtained from self, white × white and white × pink flower crosses made in the glasshouse. There were also no significant differences in the number of flowers produced by the genotypes used in the study. The observed higher frequency of white × white flower matings compared with white × pink flower matings appeared to be due to the constancy of flower colour exhibited by the butterfly pollinators Pachliopta hector and Catopsilia pyranthae during their flower visits. Observations made on the occurrence of natural self-pollination revealed that automatic self-pollination did not occur in periwinkle.     

Inheritance of resistance to Pythium dieback in the medicinal plant periwinkle

Inheritance of resistance to dieback, caused by Pythium aphanider‐matum, in periwinkle (Catharanthus roseus) was studied using a dieback‐resistant dwarf mutant, Dw, with a green stem, and a susceptible accession, OR, with a purple stem. Plants of parental F₁, F₂ and backcross generations were evaluated for resistance to dieback in a randomized complete block design with four replications, in a field plot that had been continuously planted with periwinkle in previous years and where dieback severity was high. Resistance to dieback was evaluated on two dates, i.e. when distinct differences between plants of the parental strains were noticed. From both quantitative and qualitative analyses of data, resistance to dieback appeared to be governed by a single gene (with a broad‐sense heritability of 0.85 and 0.79, depending on the date of evaluation), and was inherited independently of genes governing dwarfness and stem pigmentation.     

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.     

Characteristics and Inheritance of Seed-Ageing Induced Mutations in Lettuce (Lactuca sativa L.)

Mutations affecting qualitative traits were induced by seed ageing in lettuce. The mutant plants were isolated in the A₂ generation and included chlorophyll-deficient types (chlorotica, lutescens, chlorina-virescens, luteo and viridalbo maculate), and morphological variants (dwarf and narrow, thick and curly leaf types). The leaf mutants were found to be either partially or completely sterile. Segregation pattern of the mutants in A₃ generation showed that, except for the maculata types, all chlorophyll deficiencies and the dwarf mutant are controlled by single recessive nuclear genes. The genetic status of the leaf mutants was not clear, due to possible pleiotropic effect of the mutant genes in inducing gametophytic sterility. The maculata mutants exhibited sorting out of the normal and chlorophyll deficient regions during vegetative development and segregated for different degrees of chlorophyll deficiency in selfed progenies. The maculata mutants probably originated by plastome mutations induced by nuclear mutator genes.     

Screening of Chinese cabbage mutants produced by 60Co γ‐ray mutagenesis of isolated microspore cultures

Since the release of the Chinese cabbage genome sequence, increasing interest has focused on the functional analysis of unidentified genes in Chinese cabbage. Mutant analysis forms the basis of functional genomics research. To produce a variety of Chinese cabbage mutants in the same genetic background, buds containing late uninucleate spores from a doubled haploid line of the Chinese cabbage variety ‘Fukuda 50’ were irradiated with ⁶⁰Co γ‐rays at doses of 20, 40 and 60 Gy. Then, the treated microspores were isolated and cultured. A total of 492 putative M₀ mutants were isolated from 1483 regenerated plants. Of these, six M₁ mutants were verified; the mutant frequency was 0.41%. These mutants comprise a mutant library that includes one plant shape mutant, two flower mutants and three male sterile mutants. Pollen viability detection and DNA flow cytometry were used to determine the ploidy of the regenerated plants. Some of the mutants isolated in this study may be useful for Chinese cabbage breeding and functional genomics research.     

Inheritance of plant height and allelism tests of the dwarfing genes in Chinese barley

In order to find new dwarfing genes, the inheritance of plant height in 25 Chinese barley dwarf accessions was studied and allelism tests carried out, not only between the dwarfing genes found but also with the known dwarfing genes uz, br, sdw and denso. The results showed that out of the 25 dwarf accessions, 20 were due to monogenic recessiveness and four to digenic recessiveness. Only the short plant character in ‘1974E’ was controlled by a recessive together with a dominant dwarfing gene. In the present study, seven recessive and one dominant new dwarfing genes were identified. Five recessive genes were found in the monogenic mutants ‘91G318’, ‘91D27’ and ‘93‐597’ and in the Tibetan monogenic dwarf landraces ‘Jia Jiu’ and ‘BQK’, respectively. The other two recessives were found in the Tibetan digenic dwarf landraces ‘ZLL’ and ‘ZLLQK’, and the one dominant gene in the digenic mutant ‘1974E’.     

水稻半矮秆基因iga-1的鉴定及精细定位

在前期通过空间诱变获得半矮秆隐性突变基因iga-1的基础上,进一步对iga-1进行鉴定。农艺性状调查表明携带iga-1的矮秆株系CHA-2、CHA-2N与原种特籼占13相比存在明显变异。节间长度测量显示CHA-2、CHA-2N节间比例正常,属dn型。外源GA₃处理、内源GA₃测定和α-淀粉酶活性检测揭示iga-1与GA₃调控无关。利用CHA-2与粳稻品种02428杂交获得的F₂群体将iga-1定位在水稻第5染色体两个InDel标记DL18和DL19间32.01 kb的物理距离内。该区域有5个阅读框架,其中包括赤霉素信号传导调控基因D1。序列分析表明CHA-2、CHA-2N和特籼占13在D1位点上基因组序列不存在差异,推测D1并非iga-1的候选基因。比较水稻第5染色体上其他矮秆基因发现iga-1可能与半矮秆基因sd-7来自同一位点。     

Genetic mapping of the barley lodging resistance locus Erectoides‐k

The barley (Hordeum vulgare L.) mutant erectoides‐k.32 (ert‐k.32) was isolated in 1947 from an X‐ray‐mutant population of cultivar ‘Bonus’. The mutant was released as a cultivar in 1958 with the name ‘Pallas’ – one of the first cereal crop cultivars developed from induced mutants. ‘Pallas’ is a semi‐dwarf barley cultivar known for its culm stability and resistance to lodging. In total, eight allelic ert‐k mutants are known that show different phenotypic strength concerning culm length and spike architecture. They represent alternatives to the widely used, but pleiotropic ‘Green Revolution’ alleles of the Sdw1 (semidwarf1/denso) and Uzu1 (semi‐brachytic1) genes in breeding of robust elite barley cultivars. In the present study, we locate Ert‐k to a 15.7‐cM region in the centromeric region of chromosome 6H. Although the interval is estimated to contain approximately 700 genes, the work provides a solid foundation for the identification of the underlying mutations causing the ert‐k lodging‐resistant phenotype. In addition, the linked markers could be used to follow the ert‐k mutant genotype in marker‐assisted selection of new lodging‐resistant barley cultivars.     

Foliar disease resistant and productive mutants from the introgression lines of peanut (Arachis hypogaea)

With an objective of developing the induced mutants for superior productivity and resistance to foliar diseases in peanut, two introgression lines (IL-3 and IL-4) derived from ABK genomes of peanut were subjected to gamma and sodium azide mutagenesis. Evaluation of M₁, M₂ and M₃ generations could identify foliar disease resistant and productive mutants. Large-scale evaluation of M₄ mutants during the rainy season of 2017 confirmed the superiority of 12 mutants over the respective parents and the best check (GPBD 4). The selected mutants were resistant to leaf spots (early and late) and rust diseases, and significantly more productive than the parents and GPBD 4. ddRAD-Sequencing of these mutants identified the single nucleotide polymorphisms (SNPs) when compared to their parents. The genes harbouring these structural mutations were also identified. However, no copy number variations (CNVs) were observed between the mutants and the parents. The selected mutants carried resistant-type alleles at the marker loci linked to foliar disease resistance. These promising mutants (M₈) are currently under initial trials of variety development for commercial release.     

Inheritance and potentials of a mutated dwarfing gene ndf1 in Brassica napus

A dwarf mutant ‘NDF‐1′, approximately 70 cm high, was derived from a 200‐cm high doubled haploid (DH) line ‘3529’ (Brassica napus), seeds of which were jointly treated with chemical inducers and bombardment of fast neutron. The leaves of the ‘NDF‐1’ mutant were wrinkled and thicker compared with the wild‐type control. The mutant had much lower values than its original parents for all agronomic traits, except for its seed weight. A genetic analysis revealed that dwarfism is under the control of a major gene (designated as ndf1) with a mainly additive effect and non‐significant dominance effect. Because of the high level of resistance to lodging, breeding programmes for double low dwarf oilseed rape and heterosis utilization were initiated. Some new dwarf strains with improved agronomic performance were developed. The hybrid of the cross between the tall parent and the dwarf line showed increased harvest index and significantly higher seed yield than the tall parent or the control variety ‘Zhongyou 821’ and presented an estimated heterosis vigour rate as high as 12.5–25.8%. The dwarf trait will be a promising marker for a simple, economic and efficient way to control the purity of F₁ hybrid varieties in hybrid production of B. napus.     

Fine mapping of a day‐length‐sensitive dwarf gene in rice

Dwarf mutants are valuable and crucial resources for genetic research and crop breeding programme in rice. In this study, we identified a dwarf mutant derive from tissue culture, which exhibited a delayed heading date and dwarfism under long‐day growth conditions, suggesting the heading date of dwarf mutant was sensitive to day length. Based on 2000 F₂ mutant‐like individuals from the cross of the mutant and a Japonica var. ‘IRAT129’, the dwarf gene was finally narrowed into a 512‐kb region near the centromere on chromosome 9. According to the sequence analysis of a delimited region, 21 genes had base alternations either in promoters (15 SNPs) or in coding regions (6 InDels) among 73 annotated genes, and five genes were confirmed sequence alternations resulting from their expression mainly in the vegetative organs. Given to the RNAi plants of the five genes incapable to mimic dwarf and late heading date phenotype, the candidate gene remains to be identified by other genetic or molecular methods. Therefore, all these results give us informative foundation for the day‐length‐sensitive dwarf gene isolation.     

Inheritance of plant elongation ability at seedling stage in rice

Summary In a comparison of methods to study inheritance of plant elongation ability, 15-, 20-, 25-, 30-, and 35-day-old F₂ populations of a cross between Baisbish (floating variety) and IR42 (nonelongating semidwarf modern variety) of rice, (Oryza sativa L.) were subjected to 65 cm water depth for 7 days. Frequency distribution of plant height before and after submergence was obtained. Bimodal curves in 15-, 20- and 25-day-old populations gave good fits to 9:7 elongating: nonelongating plants, suggesting that elongation was due to two dominant complementary genes. Segregation in the 30-day-old population was not clear-cut. A seedling age of 20 days was subsequently chosen for further studies.Two F₂'s involving floating rice and a nonelongating semidwarf; four F₂'s involving floating rice and an elongating semidwarf; and two F₂'s involving elongating and nonelongating semidwarf parents were studied with 20-day-old seedlings in the same way. Floating rice combinations with nonelongating semidwarf parents as well as with elongating semidwarf parents segregated into 9:7 elongating: nonelongating ratio. It is possible that because elongating and nonelongating dwarf parents did not differ much in elongation ability at seedling age, their combination with floating rice parents provided similar segregation. The F₂ distributions for height in elongating and nonelongating dwarf cross combinations were continuous with one peak.Genetic constitution of parents proposed are Sd1 Sd1 El El for floating parents, sd1 sd1 El El for semidwarf elongating, and sd1 sd1 el el for dwarf nonelongating.     

Evaluation of breeding semidwarf rice by induced mutation and hybridization

Summary Semidwarf indica varieties of rice (Oryza sativa L.) were developed by induced mutation. They generally have a yield capacity equal to that of the semidwarf varieties (T(N)1, IR 8 and others) selected from hybridization. Genetic studies showed that semidwarf genes induced in the mutants and spontaneous ones in existing varieties are at the same locus. These semidwarf mutants, selected in Taiwan and from an American long-grain variety, Centura Patna 231, were compared with their parents, the semidwarf variety Dee-goe-woo-gen, T(N)1 and IR 8. The semidwarf mutants average about 35% shorter than their tall parents. The plant height is reduced by shortening five internodes from the top, while panicle length is not affected. The mutants, Dee-geo-woo-gen, and improved short-statured varieties had similar patterns of internode elongation. The mutants derived from varieties sensitive to photoperiod have lost the sensitivity. One more crop can be grown within a year. All induced and hybrid-derived semidwarf varieties were highly susceptible to bacterial leaf blight when Xanthomonas oryzae was inoculated although some of their parents were moderately resistant. The mutation rate from irradiation of this semidwarf gene was estimated to be 0.1% in I-kung-bau, a native variety of Taiwan. This particular type of mutation has not yet been reported in japonica varieties. However, several promising strains with reduced plant height, earlier maturity and more grain yielding were recently obtained from the California japonica variety, Calrose, by X-irradiation. The merits of mutation breeding in rice are discussed.     

Potential of gamma radiation enhancing the biosynthesis of tropane alkaloids in black henbane (Hyoscyamus niger L.)

Summary A productive yellow flowered mutant with high tropane alkaloid content was isolated in the M₂ generation of gamma irradiated (Co⁶⁰ at 40 kr) progenies ofHyoscyamus niger. Mutant progenies, on an average, were capable of synthetising more than twice crude alkaloid (0.147% to 0.221%) in M₃ through M₆ generations compared to the parental control (0.065 to 0.106%). In pilot scale trial (M₇), two elite lines, Y-15 and Y-17 were found to contain 0.390% and 0.318% crude drug in contrast to 0.211% in an improved inbred used as control. However, owing to conducive plant morphology, Y-17 registered high yield of dry biomass, hence gave more than double yield of crude drug than the improved control (inbred) (23 kg/ha against 11 kg/ha). Also being superior in hyoscyamine and hyoscine yields, it was redesignated as var.Aela, and released for commercial cultivation in India, thus signifying the efficiency of radiation breeding for crop improvement.     

小麦矮秆突变体的鉴定及其突变性状的关联分析

矮秆突变体是小麦育种和株高遗传研究的重要基因资源。通过‘云麦53’成熟种子的EMS (Ethyl methyl sulfonate)诱变及诱变植株连续自交,获得了33个M3代候选突变体。通过诱变亲本与M2和M3代候选植株的株高差异分析,筛选到26个矮秆突变体,其株高变幅为(13.61±0.11)~(44.08±1.73) cm。基于8个矮秆基因的12个特异性标记检测发现, 26个矮秆突变体至少携带2个矮秆基因标记位点。除株高外, 26个矮秆突变体还携带穗长、小穗密度、节间数和平均节间长4个不同突变性状。26个矮秆突变体可聚为5个亚类,第1亚类的小穗数和小花数最少;第2亚类的株高最矮,穗长和平均节间长最短,小穗密度最高;第3亚类突变体的节间数最少。株高与平均节间长和节间数呈极显著相关,偏相关系数分别为0.94、0.58,但与穗长、小穗数、小花数和小穗密度4个性状无相关性。26个矮秆突变体的株高与平均节间长和节间数关联遗传,携带不同的突变基因组合,可用于小麦矮化育种,以及株高、穗长和小穗密度等性状的遗传机制研究。     

Isolation of a Spontaneous Chromosome Translocation in Common Wheat

The genes Ms2 for male sterility and Rht10 for dominant dwarfing located on the short arm of chromosome 4D in common wheat arc closely linked. Male sterile, dwarf F₁ plants from the cross of male sterile‘Chinese Spring’× dwarf‘Ai-bian’were backcrossed with the variety‘Chinese Spring, From this offspring a spontaneous chromosome translocation was isolated resulting in a recombinant male sterile and dwarf genotype.     

籼稻多蘖矮半矮秆基因的遗传分析和基因定位

对籼稻标记基因系材料多蘖矮的遗传分析表明, 其矮生性状是由2对隐性半矮秆基因控制的,分别为sd1和一个新的半矮秆基因,该基因初步定名为sdt3。以多蘖矮与南京6号杂交F₂的分离群体为基础,应用SSR标记进行连锁分析,将半矮秆基因sdt3定位于第11染色体的SSR标记SSR98和SSR35之间,分别相距0.06 cM、0.13 cM,二者之间的物理距离约为93kb。以南京6号为轮回亲本与多蘖矮进行回交和自交获得由半矮秆基因sdt3控制的近等基因系（新多蘖矮）,以赤霉素处理表明由sdt3控制的半矮秆系新多蘖矮对赤霉素不敏感。     

Semidwarf gene sdk has pleiotropic effects on rice (Oryza sativa L.) plant architecture

Ideal plant architecture has become a significant objective in high‐yield rice breeding. In this study, a new semidwarf gene in sdk, a spontaneous dwarf mutant of ‘Kasalath’, was characterized and genetically mapped. The mutant has smaller panicles and seeds, reduced awns, and is 6–8 days earlier in heading than ‘Kasalath’. There was no significant difference in number (No.) of seeds and fertile seeds per panicle. Genetic analysis indicated that the dwarf character of sdk is controlled by a recessive gene, semidwarf k (sdk). From a segregating population of about 65 000 individuals, 4987 sdk‐type individuals were sampled. Gene sdk was located in a genomic region of 2622 kb flanked by markers R‐12 and R‐43 in the centromeric region of chromosome 2. The results provide an opportunity for gaining an understanding of the molecular mechanisms underlying sdk and opening possibilities for its use in rice breeding programmes.