Extension of thein vivohaploid induction system from maize to wheat

AbstractDoubled haploid breeding technology has been one of the most important techniques for accelerating crop breeding. In compare toin vivohaploid induction in maize, which is efficient and background independent, wheat haploid production by interspecific hybridization pollinated with maize is influenced by genetic background and requires rescue of young embryos. Here, we analyzed the homologues of maize haploid induction geneMTL/ZmPLA1/NLDin several crop species systematically, the homologues are highly conserved in sorghum, millet and wheat etc. Since wheat is a very important polyploidy crop, as a proof of concept, we demonstrated that thein vivohaploid induction method could be extended from diploid maize to hexaploid wheat by knocking out the wheat homologues (TaPLAs). Result showed that double knock-out mutation could trigger wheat haploid induction at ~ 2%-3%, accompanied by 30% - 60% seed setting rate. The performance of haploid wheat individual showed shorter plant, narrower leaves and male sterile. Our results also revealed that knockout ofTaPLA-A andTaPLA-D do not affect pollen viability. This study not only confirmed the function of the induction gene and explored a new approach for haploid production in wheat, but also provided an example that thein vivohaploid induction could be applied in more crop species with different ploidy levels. Furthermore, by combining with gene editing, it would be a fast and powerful platform for traits improvement in polyploidy crops breeding.

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Optimization of chromosome doubling treatment for efficient in vivo doubled haploid production in maize

Israel Journal of Plant Sciences ◽

10.1163/22238980-bja10020 ◽

2021 ◽

pp. 1-10

Author(s):

Sourbh Kumar ◽

Uttam Chandel ◽

Satish Kumar Guleria

Keyword(s):

Doubled Haploid ◽

Seed Set ◽

Chromosome Doubling ◽

Pollen Viability ◽

Haploid Inducer ◽

Haploid Production ◽

Maize Genotypes ◽

Different Doses

Abstract An investigation to optimize the protocol for application of colchicine for enhancing the doubled haploid production in maize was done. 106 maize genotypes were used as maternal parents, whereas, pollen source involved tropically adopted haploid inducer (TAIL P1 and TAIL hybrid). After the elimination of chromosomes of inducer lines, haploid seeds were obtained from the crosses. Haploid seedlings were treated with three different doses, such as 0.04, 0.06 and 0.08 per cent of colchicines for different durations (8, 12 and 15 hours). The response of various colchicine concentrations applied for different time durations revealed significant differences at P ≤ 0.05 for various parameters viz., per cent plants survivability, stalk colour, the fertility of tassel, silk present/absent, pollen viability, seed set and per cent doubled haploid formation. In maize, colchicine doses of 0.04 per cent for 12 hours and 0.06 per cent for 8 hours, respectively were established as optimum for enhanced doubled haploid production. But among these two, 0.04 per cent for 12 hours was observed to be best dose for doubled haploid production in maize.

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Pollen–pistil interaction, pistil histology and seed production in A×B grain sorghum crosses under chilling field temperatures

The Journal of Agricultural Science ◽

10.1017/s0021859609990396 ◽

2009 ◽

Vol 148 (1) ◽

pp. 73-82 ◽

Cited By ~ 14

Author(s):

M. E. CISNEROS-LÓPEZ ◽

L. E. MENDOZA-ONOFRE ◽

H. A. ZAVALETA-MANCERA ◽

V. A. GONZÁLEZ-HERNÁNDEZ ◽

G. MORA-AGUILERA ◽

...

Keyword(s):

Pollen Tube ◽

Seed Production ◽

Pollen Tube Growth ◽

Pollen Germination ◽

Vascular Tissue ◽

Pollen Viability ◽

Tube Growth ◽

Male Sterile ◽

Pollen Diameter

SUMMARYSix pairs of isogenic lines of sorghum (Sorghum bicolor L. Moench) were sown in field plots in Montecillo, State of México (2240 m altitude), in 2005 and 2006. Crosses A (♀)×B (♂) were done in each pair. In A-lines, the length of pistil, stigma, style and ovary, as well as the ovary width, were measured. In B-lines, pollen diameter, viability (cytoplasm density) and production were evaluated. Pollen germination and pollen tube growth in the pistils of the A-lines, were quantified in vivo with aniline blue and epifluorescence 18 h after pollination (HAP), while fertilized pistils were counted at 96 HAP. Histological studies on both pollinated and non-pollinated pistils were performed in one male-sterile line. Seed yield, mean-seed weight, seeds per panicle and seed set (SS; seeds/flower/panicle) were determined at harvest. Pollen viability was the variable most related to pollen germination and pollen tube growth. Stigma receptivity was not associated with its morphology. Growth of the pollen tube in stigma, style and ovary was observed in the transmitting tissue 18 HAP, running parallel to the vascular tissue. Yield under chilling field temperatures (minimum average of 6 and 8°C) prevailing during flower development and pollination ranged from 7 to 12 g/panicle. The differences in seed production and SS among A×B crosses did not depend on the amount and viability of pollen.

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A genotype independent DMP-HI system in dicot crops

10.1101/2021.06.21.449224 ◽

2021 ◽

Author(s):

Yu Zhong ◽

Baojian Chen ◽

Dong Wang ◽

Xijian Zhu ◽

Yuwen Wang ◽

...

Keyword(s):

Doubled Haploid ◽

Haploid Induction ◽

Crop Breeding ◽

Single Generation ◽

The Way

Doubled haploid (DH) technology is used to obtain homozygous lines in a single generation, which significantly accelerates the crop breeding trajectory. Traditionally, in vitro culture is used to generate DHs, but is limited by species and genotype recalcitrance. In vivo haploid induction (HI) through seed is been widely and efficiently used in maize and was recently extended to several monocot crops. However, a similar generic and efficient HI system is still lacking in dicot crops. Here we show that genotype-independent in vivo HI can be triggered by mutation of DMP genes in tomato, rapeseed and tobacco with HI rates of ~1.9%, 2.4% and 1.2%, respectively. The DMP-HI system offers a robust DH technology to facilitate variety improvement in these crops. The success of this approach and the conservation of DMP genes paves the way for a generic and efficient genotype-independent HI system in other dicot crops.

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Doubled haploid production in maize under Sub-montane Himalayan conditions using R1-nj-based haploid inducer TAILP1

Indian Journal of Genetics and Plant Breeding (The) ◽

10.31742/ijgpb.80.3.4 ◽

2020 ◽

Vol 80 (03) ◽

Author(s):

R. K Khulbe ◽

A. Pattanayak ◽

Lakshmi Kant ◽

G. S. Bisht ◽

M. C. Pant ◽

...

Keyword(s):

Doubled Haploid ◽

Sweet Corn ◽

Haploid Induction ◽

Maize Breeding ◽

Line Production ◽

Haploid Inducer ◽

Breeding Programmes ◽

Source Populations ◽

Developmental Phases

The use of in vivo haploid induction system makes the doubled haploid (DH) technology easier to adopt for the conventional maize breeders. However, despite having played an important role in the initial developmental phases of DH technology, Indian maize research has yet to harvest its benefits. Haploid Inducer Lines (HILs) developed by CIMMYT are being widely used in maize breeding programmes in many countries including India. There, however, is no published information on the efficiency of DH line production using CIMMYT HILs in Indian maize breeding programmes. In the present study, the efficiency of DH production using CIMMYT’s tropically adapted inducer line TAILP1 was investigated with eight source populations including two of sweet corn. The average haploid induction rate (HIR) of TAILP1 was 5.48% with a range of 2.01 to 10.03%. Efficiency of DH production ranged from 0.14 to 1.87% for different source populations with an average of 1.07%. The information generated will be useful for maize breeders intending to use DH technology for accelerated development of completely homozygous lines.

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Knock out of tmem38b by CRISPR/Cas9 in zebrafish unveils the in vivo role of Trimeric intracellular cation (TRIC) channel B on cell homeostasis

Bone Reports ◽

10.1016/j.bonr.2021.101054 ◽

2021 ◽

Vol 14 ◽

pp. 101054

Author(s):

Laura Leoni ◽

Valentina Daponte ◽

Francesca Tonelli ◽

Roberta Gioia ◽

Silvia Cotti ◽

...

Keyword(s):

Cell Homeostasis ◽

Knock Out

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Cosegregation of Single Genes Associated with Fertility Restoration and Transcript Processing of Sorghum Mitochondrial orf107 and urf209

Genetics ◽

10.1093/genetics/150.1.383 ◽

1998 ◽

Vol 150 (1) ◽

pp. 383-391 ◽

Cited By ~ 1

Author(s):

Hoang V Tang ◽

Ruying Chang ◽

Daryl R Pring

Keyword(s):

Fertility Restoration ◽

Mitochondrial Gene ◽

Pollen Viability ◽

Single Gene ◽

Dominant Gene ◽

Nuclear Gene ◽

Male Sterile ◽

Reading Frame ◽

Transcript Processing ◽

Backcross Populations

Abstract Defective nuclear-cytoplasmic interactions leading to aberrant microgametogenesis in sorghum carrying the IS1112C male-sterile cytoplasm occur very late in pollen maturation. Amelioration of this condition, the restoration of pollen viability, involves a novel two-gene gametophytic system, wherein genes designated Rf3 and Rf4 are required for viability of individual gametes. Rf3 is tightly linked to, or represents, a single gene that regulates a transcript processing activity that cleaves transcriptsof orf107, a chimeric mitochondrial open reading frame specific to IS1112C. The mitochondrial gene urf 209 is also subject to nucleus-specific enhanced transcript processing, 5′ to the gene, conferred by a single dominant gene designated Mmt1. Examinations of transcript patterns in F2 and two backcross populations indicated cosegregation of the augmented orf107 and urf209 processing activities in IS1112C. Several sorghum lines that do not restore fertility or confer orf107 transcript processing do exhibit urf209 transcript processing, indicating that the activities are distinguishable. We conclude that the nuclear gene(s) conferring enhanced orf107 and urf209 processing activities are tightly linked in IS1112C. Alternatively, the similarity in apparent regulatory action of the genes may indicate allelic differences wherein the IS1112C Rf3 allele may differ from alleles of maintainer lines by the capability to regulate both orf107 and urf209 processing activities.

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Lack of WWC2 Protein Leads to Aberrant Angiogenesis in Postnatal Mice

International Journal of Molecular Sciences ◽

10.3390/ijms22105321 ◽

2021 ◽

Vol 22 (10) ◽

pp. 5321

Author(s):

Viktoria Constanze Brücher ◽

Charlotte Egbring ◽

Tanja Plagemann ◽

Pavel I. Nedvetsky ◽

Verena Höffken ◽

...

Keyword(s):

Signalling Pathway ◽

Control Group ◽

Knock Out ◽

Ocular Angiogenesis ◽

Sprouting Angiogenesis ◽

Adult Mice ◽

Upstream Regulator ◽

Knock Out Mice ◽

Hippo Signalling

The WWC protein family is an upstream regulator of the Hippo signalling pathway that is involved in many cellular processes. We examined the effect of an endothelium-specific WWC1 and/or WWC2 knock-out on ocular angiogenesis. Knock-outs were induced in C57BL/6 mice at the age of one day (P1) and evaluated at P6 (postnatal mice) or induced at the age of five weeks and evaluated at three months of age (adult mice). We analysed morphology of retinal vasculature in retinal flat mounts. In addition, in vivo imaging and functional testing by electroretinography were performed in adult mice. Adult WWC1/2 double knock-out mice differed neither functionally nor morphologically from the control group. In contrast, the retinas of the postnatal WWC knock-out mice showed a hyperproliferative phenotype with significantly enlarged areas of sprouting angiogenesis and a higher number of tip cells. The branching and end points in the peripheral plexus were significantly increased compared to the control group. The deletion of the WWC2 gene was decisive for these effects; while knocking out WWC1 showed no significant differences. The results hint strongly that WWC2 is an essential regulator of ocular angiogenesis in mice. As an activator of the Hippo signalling pathway, it prevents excessive proliferation during physiological angiogenesis. In adult animals, WWC proteins do not seem to be important for the maintenance of the mature vascular plexus.

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The Extracellular Small Leucine-Rich Proteoglycan Biglycan Is a Key Player in Gastric Cancer Aggressiveness

Cancers ◽

10.3390/cancers13061330 ◽

2021 ◽

Vol 13 (6) ◽

pp. 1330

Author(s):

Filipe Pinto ◽

Liliana Santos-Ferreira ◽

Marta T. Pinto ◽

Catarina Gomes ◽

Celso A. Reis

Keyword(s):

Clinical Value ◽

Knock Out ◽

Angiogenic Potential ◽

In Vivo Experiments ◽

Cancer Aggressiveness ◽

Advanced Stages ◽

Oncogenic Gene

Biglycan (BGN gene), an extracellular proteoglycan, has been described to be associated with cancer aggressiveness. The purpose of this study was to clarify the clinical value of biglycan as a biomarker in multiple independent GC cohorts and determine the in vitro and in vivo role of biglycan in GC malignant features. We found that BGN is commonly over-expressed in all analyzed cohorts, being associated with disease relapse and poor prognosis in patients with advanced stages of disease. In vitro and in vivo experiments demonstrated that biglycan knock-out GC cells display major phenotypic changes with a lower cell survival, migration, and angiogenic potential when compared with biglycan expressing cells. Biglycan KO GC cells present increased levels of PARP1 and caspase-3 cleavage and a decreased expression of mesenchymal markers. Importantly, biglycan deficient GC cells that were supplemented with exogenous biglycan were able to restore biological features, such as survival, clonogenic and migratory capacities. Our in vitro and in vivo findings were validated in human GC samples, where BGN expression was associated with several oncogenic gene signatures that were associated with apoptosis, cell migration, invasion, and angiogenesis. This study provided new insights on biglycan role in GC that should be taken in consideration as a key cellular regulator with major impact in tumor progression and patients’ clinical outcome.

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Neuropilin-1 on hematopoietic cells as a source of vascular development

Blood ◽

10.1182/blood-2002-01-0119 ◽

2003 ◽

Vol 101 (5) ◽

pp. 1801-1809 ◽

Cited By ~ 35

Author(s):

Yoshihiro Yamada ◽

Yuichi Oike ◽

Hisao Ogawa ◽

Yasuhiro Ito ◽

Hajime Fujisawa ◽

...

Keyword(s):

Endothelial Cells ◽

Fetal Liver ◽

Vascular Development ◽

Hematopoietic Cells ◽

Vegf Receptor ◽

Knock Out ◽

Neuropilin 1 ◽

Vasculogenesis And Angiogenesis

Neuropilin-1 (NP-1) is a receptor for vascular endothelial growth factor-165 (VEGF165) and acts as a coreceptor that enhances the function of VEGF165 through VEGF receptor-2 (VEGFR-2). Studies using transgenic and knock-out mice of NP-1 indicated that this molecule is important for vascular development as well as neuronal development. We recently reported that clustered soluble NP-1 phosphorylates VEGFR-2 on endothelial cells with a low dose of VEGF165 and rescues the defective vascularity of the NP-1−/− embryo in vitro and in vivo. Here we show that NP-1 is expressed by CD45+ hematopoietic cells in the fetal liver, can bind VEGF165, and phosphorylates VEGFR-2 on endothelial cells. CD45+NP-1+ cells rescued the defective vasculogenesis and angiogenesis in the NP-1−/− P-Sp (para-aortic splanchnopleural mesodermal region) culture, although CD45+NP-1− cells did not. Moreover, CD45+NP-1+ cells together with VEGF165 induced angiogenesis in an in vivo Matrigel assay and cornea neovascularization assay. The extracellular domain of NP-1 consists of “a,” “b,” and “c” domains, and it is known that the “a” and “c” domains are necessary for dimerization of NP-1. We found that both the “a” and “c” domains are essential for such rescue of defective vascularities in the NP-1 mutant. These results suggest that NP-1 enhances vasculogenesis and angiogenesis exogenously and that dimerization of NP-1 is important for enhancing vascular development. In NP-1−/− embryos, vascular sprouting is impaired at the central nervous system (CNS) and pericardium where VEGF is not abundant, indicating that NP-1–expressing cells are required for normal vascular development.

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FcγRIII (CD16)-Deficient Mice Show IgG Isotype-Dependent Protection to Experimental Autoimmune Hemolytic Anemia

Blood ◽

10.1182/blood.v92.11.3997 ◽

1998 ◽

Vol 92 (11) ◽

pp. 3997-4002 ◽

Cited By ~ 92

Author(s):

Dirk Meyer ◽

Carsten Schiller ◽

Jürgen Westermann ◽

Shozo Izui ◽

Wouter L. W. Hazenbos ◽

...

Keyword(s):

Hemolytic Anemia ◽

Autoimmune Hemolytic Anemia ◽

Knock Out ◽

Effector Cells ◽

Igg Isotypes ◽

Knock Out Mice ◽

Deficient Mice ◽

Experimental Autoimmune

Abstract In autoimmune hemolytic anemia (AIHA), there is accumulating evidence for an involvement of FcγR expressed by phagocytic effector cells, but demonstration of a causal relationship between individual FcγRs and IgG isotypes for disease development is lacking. Although the relevance of IgG isotypes to human AIHA is limited, we could show a clear IgG isotype dependency in murine AIHA using pathogenic IgG1 (105-2H) and IgG2a (34-3C) autoreactive anti–red blood cell antibodies in mice defective for FcγRIII, and comparing the clinical outcome to those in wild-type mice. FcγRIII-deficient mice were completely resistent to the pathogenic effects of 105-2H monoclonal antibody, as shown by a lack of IgG1-mediated erythrophagocytosis in vitro and in vivo. In addition, the IgG2a response by 34-3C induced a less severe but persistent AIHA in FcγRIII knock-out mice, as documented by a decrease in hematocrit. Blocking studies indicated that the residual anemic phenotype induced by 34-3C in the absence of FcγRIII reflects an activation of FcγRI that is normally coexpressed with FcγRIII on macrophages. Together these results show that the pathogenesis of AIHA through IgG1-dependent erythrophagocytosis is exclusively mediated by FcγRIII and further suggest that FcγRI, in addition to FcγRIII, contributes to this autoimmune disease when other IgG isotypes such as IgG2a are involved.

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