What are the possible reasons for non-transmission of a mutation to the next generation??

Thanks to all for answers,

Dear Katie A Burnette,

Initially I was also thinking of some contamination or mosaic mutation. But my two different lines behaved the same {In another line I analysed 20 plants, found 19 (-3,-3) homozygous, also got one plant (-3,-1) hetero-zygous). This shows that -1 mutation was not vegetative. Further I think that only pollens may not be the cause because I should have at-least half of the expected number of heterozygous. Your point of female lethally is very interesting. Now the problem is that we did not store the heterozygous plants. We have only seeds from them and it seems hard to find any heterozygous among them.

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Yuan-Yeu Yau

Dear Aziz Ul Ikram,

I don't understand of your description of "heterozygous plants with different mutations combination like (-3,-1) and (-6,-2)". Could you elaborate it?

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Aziz Ul Ikram

Jiangsu University

Dear Yuan-Yeu Yau ,

Different mutations combination like (-3,-1) and (-6,-2) means, in one case one allele has 3 bases deletion and other allele has 1 base deletion. In other line one allele has 6 bases deletion and the other has 2 bases deletion. I hope this makes sense to you..

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Yuan-Yeu Yau

Hi Aziz Ul Ikram,

Thanks for the elabration. Yes, now I understand. Your mutation occurred on both alleles of the gene (biallelic gene mutation). But, I don't understand is that: if (-1, -1) can be lethal, why not (-3, -3)? Both of these mutation would create gene sequence frame-shift and should lost its function, right? Did you use a single sgRNA in the experiment? Where is sgRNA-recognizing location in the gene? In the middle of the gene?

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Aziz Ul Ikram

Jiangsu University

Dear Yuan-Yeu Yau

(-3, -3) deletion does not cause any frame-shift. It only causes the deletion of one amino acid, and in most cases it is not harmful...

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Yuan-Yeu Yau

Ok, Aziz, I got you...

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Yuan-Yeu Yau

@ Aziz Ul Ikram,

You could be dealing with a 'dominant lethal gene'. One example is described from the attached article (partial):

"A dominant lethal gene is lethal when present as a single copy per cell, even when a wild-type allele of the same gene is also present in the diploid cells. Such lethal genes are rarely studied because they cannot be propagated by breeding. Recessive lethal genes are lethal only when they are present in the homozygous or hemizygous condition. Individuals that are heterozygous for the lethal gene are viable because the wild-type allele is dominant to the lethal allele. A recessive lethal gene can be maintained and propagated in a heterozygous stock." (from the article)

So, in your case, suppose that you crate a mutation with (-3, -1) deletions for the gene, the '-3' allele is still functional (= wild type, or partially functional) and the '-1' allele is 'dominant lethal allele', then you would not get a plant with genotype of (-3. -1) for the gene. Probably you are not going to tell us the gene here, but check the details of your gene.

Dominant lethal alle
le.pdf
363.51 KB

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Yuan-Yeu Yau

@ Aziz Ul Ikram,

By the way, if your experimental results are correct, you should also look into the 'dosage effect' on your gene-of-interest. Look like your gene needs 2 wild-type copies (alleles) to make plant survive. With 0 or 1 copy of the wild-type allele, the plant will not survive. But, to draw this conculsion, more analysis need to be done to make sure that your earlier findings are legitimate.

Here is the description of 'dosage effect' from Wiki ( https://en.wikipedia.org/wiki/Gene_dosage ):

"Gene dosage is the number of copies of a particular gene present in a genome. Gene dosage is known to be related to the amount of gene product the cell is able to express, however, amount of gene product produced in a cell is more commonly dependent on regulation of gene expression. Nonetheless, changes in gene dosage (copy number variations) due to gene insertions or deletions can have significant phenotypic consequences. In eukaryotes, most genes found in the cell are expressed as autosomal genes (see autosome) and are found in two copies, alterations to this two-copy gene dosage is significantly associated with quantitative or qualitative phenotype traits and is linked to many genetic health problems such as those associated with spinal muscular atrophy and Down syndrome.[3] In Down syndrome, the gene expression on chromosome 21 has increased 50%, and this results in significant health and mental disabilities."

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