Application of Pollen Germination Media on Stigmas during Pollination Increases Seed Set in East African Highland Cooking Bananas (Musa spp.)

Round 1

Reviewer 1 Report

The presented experimental work is standard research quality. The methodology is understandable, the interpretation of the results is at a standard level, clearly demonstrating the drawn conclusions.
The discussion logically comments the results and compares them adequately with the published data. 

I have only one note on the methodology, respectively a question.

Why was the used conversion of obtained data as "predicted data (seeds)" in the work ? Are the yields of seeds so low that real data cannot be used?
This fact is not fully explained in the text.

I have no other serious comments on the submitted paper.

Author Response

Point 1: Why was the used conversion of obtained data as "predicted data (seeds)" in the work ? Are the yields of seeds so low that real data cannot be used?
This fact is not fully explained in the text.

 

Response 1: Because of the nature of banana, you cannot balance the number of bunches emerging in a given time. For example, you may get 5 bunches in March and 8 bunches in April, these bunches are most likely to be of different sizes. Our recent research (https://doi.org/10.3390/agronomy11040763) found that different bunch sizes behave differently in terms of fertility. For this reason, bunch sizes were categorized and treated as blocks or replicates.

 

Seasonality also has an effect on fertility. So to ensure that seasonality effects were captured while having a representation of all bunch sizes, a two months interval was considered. Because of the imbalances in bunch number and bunch sizes in a given two month period, data was best analysed with unbalanced design. The output of means comes as predicted means because of the regression to account for unequal numbers and say a missing bunch category. The differences were minor in comparison to just averaging. A two month period rather than say a monthly period ensured that all bunch size categories were represented. Few bunches large and small bunches were also merged to the nearest bunch size categories to avoid having many missing data.

 

I have added a sentence on lines 149 - 150 of the original document to explain predicted means. The sentence reads; Predicted means are obtain since the unbalanced design accounts for unequal numbers and missing data.

Reviewer 2 Report

I carefully reviewed the paper "Application of pollen germination media on stigmas during pollination increases seed set in East African Highland Cooking Bananas (Musa spp.)". I believe the paper is good and will be a positive contribution to the field.    

Author Response

Point 1: I carefully reviewed the paper "Application of pollen germination media on stigmas during pollination increases seed set in East African Highland Cooking Bananas (Musa spp.)". I believe the paper is good and will be a positive contribution to the field.  

 

Response 1: Thank you for this positive comment.

Reviewer 3 Report

The paper ”Application of pollen germination media on stigmas during pollination increases seed set in East African Highland Cooking Bananas (Musa spp.) by Allan Waniale et al. report that spray glucose based pollen germination media (PGM) on freshly emerged stigmas of bananas significantly increased seed set.

 Although it is important to improve the seed set of East African Highland Cooking Bananas (EAHBs) for breeding, it is question whether this paper shows the new insight to improve it.

My concerns are as follows.

 

1. what is the component of PGM? 0.25g MgSO4.7H2O…0.1g H3BO3 per liter (L111-114) is PGM? It is not clear.

 

2. I cannot understand Table 2. Table caption says that it shows seed set data after pollination with and without pollen germination media. However, there is no description of “ with and without PGM” in the table.

 

3. Table 4. It is not clear what “Mean” and “Seed” in column headings mean.

Author Response

General response 1: Improving seed set banana is such a challenge to a point that small increments are taken as big achievement. I therefore thought that banana breeding programs will find this captivating.

 

General response 2: For all sections marked "Must be improved", I have found it difficult to make the necessary changes. I expected specific comments to help me make the changes. 

 

Although it is important to improve the seed set of East African Highland Cooking Bananas (EAHBs) for breeding, it is question whether this paper shows the new insight to improve it.

My concerns are as follows.

 

Point 1: what is the component of PGM? 0.25g MgSO4.7H2O…0.1g H3BO3 per liter (L111-114) is PGM? It is not clear.

Response 1:  I used two types of media. The first type consisted of only 30 g of glucose in a liter of water. I have made this clearer by mentioning the PGM types on lines 110 and 113 of the original document.  

 

Point 2: I cannot understand Table 2. Table caption says that it shows seed set data after pollination with and without pollen germination media. However, there is no description of “with and without PGM” in the table.

 

Response 2: Pollination with and without PGM are the two pollination techniques (Pol. Tech) in the ANOVA table. I have defined this in the table legend. 

 

Point 3: Table 4. It is not clear what “Mean” and “Seed” in column headings mean.

 

Response 3: “Mean” refers to mean seed and mean pollination success. I did not want to repeat the word. I have merged the two cells and centred the word “Mean” to cover the two columns of “seed” and “pol suc”. These actions have not been marked as track changes by default.

Round 2

Reviewer 3 Report

Although the author mention that there was no significant difference in seed set per 100 fruits per bunch in ‘Enzirabahima’ …, there is no data about other cultivar. In spite of the luck data, the authors combined the data from 30 % glucose and PGM with glucose. Moreover, they discuss the PGM with or without glucose in discussion part. I don’t think it is appropriate.

Author Response

Response to Reviewer 3 Comments – Round 2

Point 1: Although the author mention that there was no significant difference in seed set per 100 fruits per bunch in ‘Enzirabahima’ …, there is no data about other cultivar. In spite of the luck data, the authors combined the data from 30 % glucose and PGM with glucose. Moreover, they discuss the PGM with or without glucose in discussion part. I don’t think it is appropriate.

 

Response 1: Pollinations were made with and without pollen germination media (PGM). Within PGM, there were two types of PGM. The first type was prepared with only 30 g/l of glucose. The second type of media was complete PGM as used for in vitro pollen germination studies.  See lines 110 – 116 of the original manuscript, also made clearer in the revised version.

 

The first comparison is for the two types of PGM, that is PGM in 2016 and PGM in 2018 – see lines 155 – 157 and table 1. Control in 2016 and 2018 was also compared to check for consistence. Since the two types of media were not significantly different (lines 162 – 168), we merged PGM data, that is PGM with only 30 g/l glucose and complete PGM. So the merged PGM data is the one compared with control technique – Pollination without PGM (lines 169 – 176).

 

There were very few bunches pollinated for Mshale in 2016, a total of 15 bunches (Table 5). Reliable comparison could not be made for PGM 30 g/l glucose and complete PGM. A sentence on the first paragraph of results has been added. It reads; Reliable comparisons could not be made for ‘Mshale’ because only 15 bunches in total were pollinated in 2016. This was so because of staggered planting (line 122 – 123). We planted on different dates because we could not obtain all planting material at once. No bunches of Nshonowa were ready for pollination (with and without PGM) in 2016 (line 111 – 112). Please see the discussion part on two media types from line 238 – 250.

 

I there think it is appropriate to compare the two pollination techniques as they were significantly different. It was the two media types that were not significant.