Sterilization Effect of Cooking Process for Guilin Rice Noodles Based on Heat Conduction Model

Guilin rice noodles, a unique cuisine from Guilin, Guangxi, is renowned both domestically and internationally as one of the top ten “Guilin Classics”. Utilizing a heat conduction model, this study explores the effectiveness of the cooking process in sterilizing Guilin rice noodles before consumption. The model assumes that a large pot is filled with boiling water which is maintained at a constant high temperature heat resource through continuous gentle heating. And the room temperature is set as the initial temperature for the preheating process and the final temperature for the cooling process. The objective is to assess whether the cooking process achieves satisfactory sterilization results. The temperature distribution function of rice noodle with time is analytically obtained using the separation of variables method in the three-dimensional cylindrical coordinate system. Meanwhile, the thermal diffusion coefficient of Guilin rice noodles is obtained in terms of Riedel’ theory. By analyzing the elimination characteristics of Pseudomonas cocovenenans subsp. farinofermentans, this study obtains the optimal time required for effective sterilization at the core of Guilin rice noodles. The results show that the potential Pseudomonas cocovenenans subsp. farinofermentans will be completely eliminated through continuously preheating more than 31 seconds during the cooking process before consumption. This study provides a valuable reference of food safety standards in the cooking process of Guilin rice noodles, particularly in ensuring the complete inactivation of potentially harmful strains such as Pseudomonas cocovenenans subsp. farinofermentans.

Observation on Flowering Habit of Reverse Genic Sterile Rice FHS

[Objective] This study was to use reverse genic male sterile FHS better in field production. [Method] Low energy nitrogen ion beam was taken as a mutation source to conduct mutagenic treatment for indica rice 99-02 and the mutant FHS with special fertility that was isolated from their offspring. Some characteristics such as flowering habit and stigma exsertion rate of FHS were observed in this experiment. [Result] The reverse genic sterile rice FHS had an obvious peak flowering stage from 10:00 to 10:30, while the second peak flowering was from August 5 to August 10. Compared with PA64S, FHS flowered early and its flowering time was concentrated, showing that it is for seed propagation. The stigma exsertion rate of FHS was 85.8% and low exsertion rate was good for the purity of seed. [Conclusion] The reverse genic sterile rice FHS had good value in use, besides, it could also be used as comparison material for studying fertility alternation mechanism of photoperiod sensitive genic male sterile rice.

Immunohistochemical Analysis of IAA in Anthers of the Photoperiod-sensitive Genic Male-sterile Rice

The immunohistochemical localization of IAA and the comparison of their relative levels were carried out for the first time in the anthers of Nongken 58S and its wild type Nongken 58 (Oryza sativa subsp. japonica) after long_day and short_day treatments. The distribution of free_IAA in anthers and its dynamic variation could be reflected by this method. The results showed that the IAA level in the anthers of Nongken 58S after long_day treatment was much lower than that in short_day_treated Nongken 58S and those in wild type Nongken 58 in five stages from pistil and stamen primordia formation to late uninucleate stage. The possible reasons for IAA deficiency in Nongken 58S_LD anthers and its relationship with fertility alteration were also discussed.

Comparative Studies on the Changes of Microtubule Distribution and Reorganization During the Meiotic Stages of Development in Normal (IR36) and a Temperature/photoperiod Sensitive Male Sterile Line (Peiai 64S) of Rice ( Oryza sativa )

Changes in the pattern of organization of microtubules in the meiotic stages of development of pollen (i.e. from pre-meiotic interphase to more or less metaphase I) of a normal (IR36) and a temperature/photoperiod sensitive male sterile line (Peiai 64S) of rice were studied using immunofluorescence confocal microscopy. In IR36, from pre-meiotic interphase to metaphase I, the pattern of microtubule distribution in the meiocytes underwent a series of changes. Some new organizational patterns of microtubules (that have not been described before) were observed during microsporogenesis, including the existence of a broad band of perinuclear microtubules at the diakinesis stage of development. The pattern of microtubule distribution in the meiocytes of the male sterile line, Peiai 64S, was quite different front that seen in IR36. In Peiai 64S, the microtubules showed abnormal patterns of distribution from pre-meiotic interphase to metaphase I. For example the broad band of perinuclear microtubules seen at diakinesis in IR36 was much disorganized and loosened in Peiai 64S. The spindles formed were also very abnormal and different from the normal spindle. The appearance of abnormal microtubule distribution in the early stages of microsporogenesis may contribute to the malformation and ultimate abortion of pollen in Peiai 64S.

Study on Seed Vigor of Rice Photo-ThermoSensitive Genic Male Sterile(PTGMS) Line with Low Temperature Storage

To explore the change rule of seed vigor and seed application technology of rice PTGMS lines with low temperature storage,Zhun S,Y58S,Feng S and Guangzhan 63-4S stored in the seed bank with the temperature of 8-10 ℃ and with the relative humidity of 55% for different years were used as the materials to study the properties of seed vigor,growth and development in the field, etc. The re- sults showed that the germination potential, germination rate, germination index and vigor index of rice PTGMS lines seeds showed downward trends as the time of low temperature storage prolonged,and all of the germination rates of the seeds within five years were over 60%. The conductivity in the soaked solution of seeds was significantly increased with the prolonged low temperature storage time. Seedling rate,root length,white root number,base stem width and seedling dry weight of seeds showed downward trends as the time of low temperature storage prolonged. There was no significant difference between two integral seedling qualities of seeds within 4-6 years in low temperature storage. Compared with the germination rate of low temperature storage seeds soaking for 6,12 and 24 h,the appropriate soak time of long time low temperature storage or low vigor seeds was 12 h. A serious natural aging phenomenon occurred in low temperature storage seeds after out of the storage. The germination rate of seeds placed under natural condition for 50 d decreased obviously. The duration from seeding to heading of Zhun S,Y58S and Feng S shortened as the time of low temperature storage prolonged, while the vari- ation of panicle duration of all PTGMS lines was not obvious. In conclusion, rice PTGMS line seeds with low temperature storage in 5 years could still be used nor- mally. However.seeds after low temperature storage should be sowed as soon as possible, and the soaking time of low vigor seeds should be shortened,and the possibility that the decreased duration from seeding to heading should also be con- sidered.

Technical Specification for Producing Hybrid Pepper Seed Using Three-line Male Sterility

This paper described the whole process of three line hybrid pepper seed production in detail, including requirement of the seed production base, parent culti- vation, field management, and specified the key operation techniques in seed pro- duction, such as parental impurity removal to preserve pure state, pollen collection, pollination and seed collecting essentials. This specification is of guiding significance for the production of hybrid pepper seed and ensuring the purity of hybrid pepper seed.

Analysis of Heterosis, Combining Ability and Heritability of Cadmium Content in Brown Rice of Three-line Indica Hybrid Rice

Five cytoplasmic male sterile （CMS） lines were used as parents in an incomplete diallet cross and six restorer lines of rice design （Nc II design）. Thirty hybrid combinations with the same growth period were selected as experimental ma- terials to investigate the heterosis, combined ability and heredity of Cd content in brown rice of indica hybrid rice. According to the results, Cd content in brown rice showed a significantly negative heterosis; the general combining ability and specific combination ability of Cd content in CMS and restorer lines both reached extremely significant level （P〈0.01）, indicating that both genetic improvement of parents and e- valuation of combinations are important to the breeding of hybrid combinations with low accumulation of Cd; the broad-sense heritabitity and narrow-sense heritability of Cd content were both relatively high with slight differences, which respectively reached 97.73% and 80.10%, indicating that Cd content in brown rice mainly de- pends on the additive action of genes; in addition, parent improvement showed bet- ter effect on the selection of early generation.

High-yielding Seed Production Techniques of Disease-resistant and High-quality Hybrid Rice Sterile Line Rong 18A

Rong 18A, bred by Crop Research Institute, Chengdu Academy of Agri- culture and Forestry Sciences is a new blast-highly resistant and high-combining ability hybrid rice sterile line. It is currently one of the male sterile lines in Sichuan Province which combine well high grain quality and rice blast resistance. More than 20 combinations of Rong 18A have passed the national and provincial variety iden- tification. Combined with the climate conditions in Chengdu, the key points of high- yielding seed production techniques for Rong18A are analyzed and summarized in this paper.

Flowering Characteristics of Japonica Rice Male Sterile Lines Adaptive to High Temperature and Summer Drought

[Objective] This study was conducted to screen japonica rice male sterile lines with good flowering characteristics under high temperature and summer drought. [Method] The flowering habits of 23 japonica male sterile lines, such as flowering peak, single-panicle flowering duration, single-plant flowering duration, en- closed glume rate, glume-opening angle, the distance between glumes, stigma exer- tion rate and sigma vitality, were compared with II-32A as a control under above weather condition. [Result] The flowering characteristics had significant difference a- mong the tested rice lines. Compared with the control, three japonica rice male ster- ile lines 35478A, 35489A and 35502A had better flowering traits, such as earlier flowering peak, longer single-panicle flowering duration, longer single-plant flowering duration, lower rate of enclosed glumes, larger glume angle and larger distance be- tween glumes. 35478A performed its flowering peak from 10：00-11：00 am, 1.5 h earlier than the control; 35489A exhibited higher stigma exertion rate, up to 76.23%; and 35502A had higher stigma vitality, up to 84.68%, which kept longer time than that of the CK. [Conclusion] The results will provide high-quality resources for the breeding of new japonica rice varieties with high-yield and high-temperature toler- ance.

Effect of High Temperature on Outcrossing Characteristics at the Fertility Sensitive Period of Photo-thermo Sensitive Genic Male Sterile (PTGMS) Rice Lines

Four sterile lines （Peiai64S, Y58S, Guangzhan 63-2S and H638S） and the restoring line R527 were used as materials. Five temperature gradients （24, 27, 30, 33 and 36 ℃ in artificial climate chamber） and the natural temperature （as control） were treated to the four sterile lines for 6 d in the fertility sensitive period of heading to flowering stage, respectively, to study the effects of temperature on physiological biochemical characteristics of young panicles and outcrossing characteristics. The results showed that the percentages of exerted stigma of Peiai 64S and Y58S were the highest at 27 ℃, which were 86.81% and 86.06%, respectively, while the percentages of exerted stigma of Guangzhan 63-2S and H638S were the highest at 24 ℃, which were 76.24% and 81.76%, respectively; the stigma viability of Peiai 64S, Y58S, Guangzhan 63-2S and H638S were the best at 24 ℃, which were 1.96, 2.12, 1.74 and 1.94, respectively; the outcrossing rates of Peiai 64S, Guangzhan 63-2S and H638S were the highest at 24 ℃, which were 58.87%, 54.22% and 50.50%, respectively, while the outcrossing rate of Y58S was the highest at 27 ℃, which was 58.96%; and the contents of peroxidase （POD） of the four sterile lines increased significantly at 33 ℃ compared with the control, while the contents of malondialdehyde （MDA） and proline increased significantly at 36 ℃ compared with the control. There were differences in temperature sensitivity between the male sterile lines, and the 24 ℃ treatment during the sensitive period was the best for the fertility sensitive period of Peiai 64S, while 27 ℃ was the best temperature for Y58S, Gangzhan 63-2S and H638S.

Mechanism of Sterility and Breeding Strategies for Photoperiod/Thermo-Sensitive Genic Male Sterile Rice

To understand the male sterility mechanism of photoperiod/thermo-sensitive genic male sterile [P（T）GMS] lines in rice, the research progress on genetics of photoperiod and/or temperature sensitive genic male sterility in rice was reviewed. A new idea was proposed to explain the sterility mechanism of P（T）GMS rice. The fertility transition from sterile to fertile is the result of cooperative regulation of major-effect sterile genes with photoperiod and/or temperature sensitive genes, but not the so-called pgms gene in P（T）GMS rice. The minor-effect genes, which exhibit accumulative effect on sterility, are the important factors for the critical temperature of sterility transition. The more minor-effect genes the sterile line holds, the lower the critical temperature of sterility transition is. The critical temperature of sterility transition will be invariable if all the minor-effect genes are homozygous. The strategies for breeding P（T）GMS rice were also proposed. The selective indices of critical photoperiod and temperature for sterility transition should be set according to varietal type and ecological region. Imposing selection pressure is a key technology for breeding P（T）GMS rice with lower critical temperature for sterility, and improving the comprehensive performance of the whole traits and combining ability is vital for breeding P（T）GMS rice lines.

Development of Novel Cytoplasmic Male Sterile Source from Dongxiang Wild Rice(Oryza rufipogon)

This study was conducted to develop and characterize a novel cytoplasmic male sterile （CMS） source which was identified from Dongxiang wild rice （Oryza rufipogon） by crossing Dongxiang wild rice as female with Zhongzao 35, an indica inbred variety, as male and continuous backcrossing with Zhongzao 35. Observation under optical microscope manifested that this novel CMS belonged to typical abortion type with less pollen compared with wild abortive type cytoplasm （CMS-WA）. Sequential planting showed that this novel CMS has complete and stable male sterility. Testcross experiment showed that all the 24 tested materials including maintainer and restorer lines of CMS-WA and Honglian type cytoplasm （CMS-HL） and other indica inbred varieties are the maintainers with complete maintaining ability, suggesting that this novel CMS has fertility restoration totally different from CMS-WA and CMS-HL and belongs to a novel type of CMS. So far, we only discovered a unique fertility restoration source for this novel CMS. Inheritance analysis showed that the fertility restoration of this CMS was governed by three pairs of independent dominant genes. Prospect for application of this novel CMS system in hybrid rice breeding was also discussed.

Plant Temperature and Its Simulation Model of Thermo-Sensitive Genic Male Sterile Rice

Plant temperature （Tp） and its relations to the microclimate of rice colony and irrigation water were studied using a thermo-sensitive genic male sterile （TGMS） rice line, Pei＇ai 64S. Significant differences in the daily change of temperature were detected between Tp and air temperature at the height of 150 cm （TA）. From 8：00 to 20：00, Tp was lower than TA, but they were similar during 21：00 to next 7：00. The maximum Tp occurred one hour earlier than the maximum TA, though they both reached the minimum at 6：00. Tp fluctuated less than TA. At the same height, during 6：00-13：00, Tp was higher than air temperature （Ta）, and Tp reached the maximum one hour earlier than Ta. During the rest time on sunny day, Tp was close to or even a little lower than Ta. On overcast day, Tp was higher than Ta in the whole day, and both maximized at the same time. In addition, Tp was regulated by solar radiation, cloudage and wind speed in daytime, and by irrigation water at night. The present study indicated that a TA of 29.6℃ was the critical point, at which Tp was increased or decreased by irrigation water. Tp and the difference between water and air temperatures showed a conic relation. Tp fluctuation was also regulated by the absorption or reflection of solar radiation by leaves during daytime and release of heat energy during nighttime. By analysis on correlation and regression simulation, two models of Tp were established.

Molecular Markers and Candidate Genes for Thermo-Sensitive Genic Male Sterile in Rice

The discovery of thermo-sensitive genic male sterility(TGMS) has led to development of a simple and highly efficient two-line breeding system. In this study, genetic analysis was conducted using three F_2 populations derived from crosses between IR68301 S, an indica TGMS rice line, and IR14632(tropical japonica), Supanburi 91062(indica) and IR67966-188-2-2-1(tropical japonica), respectively.Approximately 1:3 ratio between sterile and normal pollen of F_2 plants from the three populations revealed that TGMS is controlled by a single recessive gene. Bulked segregant analysis using simple sequence repeat(SSR) and insertion-deletion(InDel) markers were used to identify markers linked to the tms gene. The linkage analysis based on the three populations indicated that the tms locus was located on chromosome 2 covering the same area. Using IR68301S × IR14632 F_2 population, the results showed that the tms locus was located between SSR marker RM12676 and InDel marker 2gAP0050058. The genetic distance from the tms gene to these two flanking markers were 1.10 and 0.82 cM, respectively.InDel marker 2gAP004045 located between these two markers showed complete co-segregation with the TGMS phenotype. In addition, InDel marker vf0206114052 showed 2.94 cM linked to the tms gene using F_2 populations of IR68301S × Supanburi 91062. These markers are useful tool for developing new TGMS lines by marker-assisted selection. There were ten genes located between the two flanking markers RM12676 and 2gAP0050058. Using quantitative real-time PCR for expression analysis, 7 of the 10 genes showed expression in panicles, and response to temperatures. These genes could be the candidate gene controlling TGMS in IR68301S.

Characterizations and identification of the candidate gene of rice thermo-sensitive genic male sterile gene tms5 by mapping

Previous study indicated that the thermo-sensitive genic malesterile(TGMS) gene in rice was regulated by temperature.TGMS rice plays an important role in hybrid rice production,because the application of the TGMS system in two-line breeding is laborsaving,timesaving,simple,inexpensive,efficient,and eliminating the limitations of the cytoplasmic male sterility(CMS) system.'AnnongS' is the first discovered and deeply studied TGMS rice lines in China.'AnnongS-1' and 'Y58S',two derivatives of TGMS line AnnongS,were both controlled by a single recessive gene named tms5,which was genetically mapped on chromosome 2.In this study,three populations('AnnongS-1' × 'Nanjing11','Y58S' × 'Q611',and 'Y58S' × 'Guanghui122') were developed and used for the molecular fine mapping of the tms5 gene.By analyzing recombination events in the sterile individuals using a total of 125 probes covering the tms5 region,the tms5 gene was physically mapped to a 19-kb DNA fragment between two markers 4039-1 and 4039-2,which were located on the BAC clone AP004039.After the construction of the physical map between two markers 4039-1 and 4039-2,a member(ONAC023) of the NAC(NAM-ATAF-CUC-related) gene family was identified as the candidate gene of the tms5 gene.

Synergistic roles of LAX1 and FZP in the development of rice sterile lemma

Rice florets are subtended by two sterile lemmas,whose origin and biological functions have not been studied extensively.Here we demonstrate that two putative transcription factors,LAX PANICLE1(LAX1)and FRIZZY PANICLE(FZP),synergistically control the development of sterile lemmas.Both LAX1 and FZP are previously known for their roles in panicle and floret development.Disruption of either LAX1 or FZP greatly reduces the number of floret development.We generated new lax1 mutants(lax1-c)using CRISPR/Cas9 gene editing technology.In addition to the expected lax panicle phenotypes,we noticed that a significant number of spikelets of lax1-c developed elongated sterile lemmas.Moreover,our characterization of lax1-RNAi plants also revealed sterile lemma phenotypes similar to lax1-c mutants.We isolated a weak allele of fzp(fzp-14)in a genetic screen for lax1–1 enhancers.The fzp-14 lax1–1 double mutants completely eliminated flower development.Interestingly,the isolated fzp-14 produced spikelets with elongated sterile lemmas.Furthermore,fzp-14 was haploid-insufficient in the lax1–1 background whereas fzp-14 heterozygous plants were indistinguishable from wild type plants.The lax1–1 fzp-14+/−also developed elongated sterile lemma as observed in lax1-c,lax1-RNAi,and fzp-14,suggesting that LAX1 and FZP synergistically control sterile lemma development.

Molecular identification of the cytoplasmic male sterile source from Dongxiang wild rice(Oryza rufipogon Griff.)

The use of cytoplasm male sterility （CMS） is crucial for three-line hybrid seed production. Two types of CMS have been discovered from Dongxiang wild rice, namely the wild abortion type CMS （CMS-WA） and the Dongxiang wild type CMS （CMS-DW）. In this study, we show the molecular identification of the two types of CMS in Dongxiang wild rice. WA352, which conferred CMS-WA, was not detected in Dongxiang wild iice, implying Dongxiang wild rice does not carry the CMS- WA source. Further analysis of WA352 in DYIA, a CMS-DW line, by PCR amplification and sequencing, revealed two insertion-deletion polymorphisms occurred in CMS-DW compared to WA352 of CMS-WA. It was reported that WA352 was comprised of an unknown origin sequence and partial sequences of three open reading frames （ORFs）, orf284, orf224 and orf288. The 42-bp insertion was located between the two segments of orf224 and orf288, which created a new chimeric ORF, orf216. This new ORF was also detected in CMS-HL. Based on the 9-bp deletion in orf284, a specific mitochon- drial marker of DW-M1 was developed, which could be used to specifically distinguish the DW-type source. Moreover, semi-quantitative RT-PCR analysis preliminarily suggested that both orf216 and orf284 could be considered as candidates for CMS-DW. These findings present a preliminary understanding of CMS-DW at the molecular level.

Inheritance of the Pale—Green Leaf Marker and Sterility Trait in Photo—Thermo Sensitive Genic Male Sterile Rice

P/TGMS (photo-thermo sensitive genie male sterility) lines with pale-green leaf color have been developed in japonica rice. The marker trait is used as an assistant selection in the production of the two-lines system hybrid rice for the improvement of F, seed purity. A joint inheritance study of both leaf color and male sterility is presented for P/TGMS line with pale-green leaf color. The segregation ratios for leaf color in the F2 populations of the three crosses showed 13 : 3 and 15 : 1 at early and late sowing stages (April 26 and June 23) respectively, implying that the leaf color is controlled by two genes with fertility gene as dominant. Sterility level is higher in the early sowing stage than that in the late sowing. The inducement of male sterility is closely related to longer day-length and higher temperature at the developmental stages of young panicle. The genes to govern the leaf color and male fertility are inherited independently.

Characterization and fine mapping of RTMS10, a semi-dominant reverse thermo-sensitive genic male sterile locus in rice

The discovery and application of environment-sensitive genic male sterile(EGMS) rice germplasm provide an easy method for hybrid rice breeding and have made great contributions to hybrid rice production. Typically, the photoperiod-and thermosensitive GMS(P/TGMS) lines utilized in two-line hybrid systems are male sterile under long day or/and high temperature but fertile under short day or/and low temperature conditions. However, Yannong S(Yn S), a reverse TGMS(rTGMS) line, is sterile under low temperature(<29℃) and fertile under high temperature(>29.5℃). Here, we report a genetic study on the rTGMS trait in Yn S. Interestingly, the F1 plants of the cross between Yn S and a cultivar, L422, were male sterile at 22℃ and completely fertile at 27℃. Moreover, the segregation ratio of fertile and sterile individuals in Yn S/L422 F2 populations changed from 1:3.05 to 2.95:1 when the ambient temperature increased, showing that the rTGMS trait exhibits semidominance in Yn S. We further found a locus on chromosome 10, termed RTMS10, which controls the rTGMS trait in Yn S. We then finely mapped RTMS10 to a ~68 kb interval between markers ID13116 and ID1318 by Yn S/L422 BC6 F2 populations. A near iso-genic line(NIL) NL1 from the BC6 F3 generation was developed and the pollen of NL1 became abnormal from the meiosis stage under low temperature. In summary, we identified an rTGMS locus, RTMS10, and provided co-segregated markers, which could help to accelerate molecular breeding of rTGMS lines and better understand the rTGMS trait in rice.

Isolation and Analysis of the Promoter of OsRacD from PhotoperiodSensitive Genic Male Sterile Rice

By using OsRacD cDNA as probe to screen the genomic library of photoperiod sensitive genic male sterile rice line Nongken 58S, a positive clone containing 2 kb promoter and 396 bp coding region of OsRacD was obtained. Compared with the promoter of OsRacD cloned by reverse PCR from normal rice variety Nongken 58 （Nongken 58N）, the homology was 99.8%, and the different nucleotides were outside the predicted response elements in promoter, suggesting that the fertility between rice varieties Nongken 58S and Nongken 58N under the long-day conditions was not attributed to the difference in the structure of OsRacD upstream regulation sequences, but to the developmental regulation of gene differential expression.