In Himachal Pradesh, the population ranged between 245cyst/250cc soil. In Himachal Pradesh, the population of the cyst ranged from minimum to maximum. In the district, Mandi samples were collected from four different areas i.e Basunti, Rewalsar, Phulladhar, and Dhanotu among these maximum populations of the cyst were collected from Basunti and Phulladhar samples. In Distt Chamba samples were collected again from four different areas i.e Ahla, Banikhet, Salooni, and Saru. Out of these four areas maximum cysts were collected from Ahla area. The third Distt was Kullu, samples were collected from four different areas; Bajaura, Bandrol, Anni, and Bhuntar. The maximum cyst population was collected from Bajaura area. From Distt Kangra samples were collected from eight different areas i.e Talpura, Kangra, Jiya, Skoh, Pariharkhar, Shahpur, Malan, and Bagoda out this Jiya area has the maximum number of the cyst. Samples were collected from Distt Una, Hamirpur, and Solan, particularly these sites i.e Akrot farm, Daulatpur, Rampur, TiraSujanpur, Nadaun, Kandaghat, and Darlaghat didn’t recover cyst.
Table 2: Showing the morphometrics of Globoderarostochiensis cyst in µm. (± is mentioned as standard deviation).
Characters
|
Measurements in µm
|
Body length
|
613.08±23.96
|
Body width
|
555.79±36.04
|
Neck length
|
148.84±38.46
|
Distance from vulval basin to anus
|
65.29 ±22.20
|
In molecular characterization after sequencing, the resulted sequence were submitted to genebank NCBHI. Sequence ID MW577347 (HPPOCH) showing 99.05% similarity with KJ636272.1 from the Netherlands. Sequence ID MZ508279 (HPPOKL) showing 95.58% similarity with KJ636272.1 from the Netherlands. Sequence ID MZ508280 (HPPOMD) showing99.05% similarity with KJ636272.1. Sequence I.D MZ518783 (HPPOPL) 98.58% similarity with KJ636272.1 from the Netherlands. All the nucleotide sequences of Himachal Pradesh showing similarities with the Netherlands.
Table 3. Population density and frequency of occurrence of PCN in various district of Himachal Pradesh in potato growing fields.
Distt
|
Location
|
Mean No. of cyst/ 250g of soil from different areas.
|
Sum
|
P.D
|
Occurrence
|
1
|
2
|
3
|
Mandi
|
Basunti
|
50
|
±1.63
|
48
|
±2.16
|
45
|
±1.63
|
143
|
47.67
|
100
|
Rewalsar
|
10
|
±0.82
|
7
|
±1.70
|
12
|
±2.45
|
29
|
9.67
|
100
|
Phulladhar
|
55
|
±`3.74
|
49
|
±5.31
|
48
|
±1.89
|
152
|
50.67
|
100
|
Dhanotu
|
5
|
±4.11
|
8
|
±4.19
|
5
|
±2.05
|
18
|
6
|
100
|
Chamba
|
Saru
|
0
|
±0.00
|
2
|
±1.63
|
0
|
±0.00
|
2
|
0.67
|
33.33
|
Ahla
|
52
|
±2.16
|
50
|
±2.45
|
54
|
±2.49
|
156
|
52
|
100
|
Banikhet
|
20
|
±5.10
|
18
|
±3.30
|
18
|
±6.48
|
56
|
18.67
|
100
|
Salooni
|
5
|
±2.05
|
4
|
±1.70
|
5
|
±3.74
|
14
|
4.67
|
100
|
Kullu
|
Bajaura
|
48
|
±5.25
|
45
|
±4.08
|
50
|
±10.96
|
143
|
47.67
|
100
|
Bandrol
|
4
|
±1.63
|
0
|
±0.00
|
0
|
±0.00
|
4
|
1.33
|
33.33
|
Anni
|
25
|
±5.10
|
16
|
±3.09
|
21
|
±4.50
|
62
|
20.67
|
100
|
Bhuntar
|
15
|
±2.45
|
14
|
±5.66
|
17
|
±2.16
|
46
|
15.33
|
100
|
Talpura
|
0
|
±0.00
|
0
|
±0.00
|
0
|
±0.00
|
0
|
0
|
0
|
Kangra
|
Kangra
|
0
|
±0.00
|
0
|
±0.00
|
0
|
±0.00
|
0
|
0
|
0
|
Jiya
|
45
|
±2.49
|
48
|
±2.49
|
0
|
±0.00
|
93
|
31
|
66.66
|
Skoh
|
0
|
±0.00
|
0
|
±0.00
|
0
|
±0.00
|
0
|
0
|
0
|
Parihar khar
|
3
|
±1.41
|
1
|
±0.82
|
0
|
±0.00
|
4
|
1.33
|
66.66
|
Shahpur
|
0
|
±0.00
|
0
|
±0.00
|
0
|
±0.00
|
0
|
0
|
0
|
Malan
|
0
|
±0.00
|
0
|
±0.00
|
0
|
±0.00
|
0
|
0
|
0
|
Bagoda
|
0
|
±0.00
|
0
|
±0.00
|
0
|
±0.00
|
0
|
0
|
0
|
Una
|
Akrot farm
|
0
|
±0.00
|
0
|
±0.00
|
0
|
±0.00
|
0
|
0
|
0
|
Daulatpur
|
0
|
±0.00
|
0
|
±0.00
|
0
|
±0.00
|
0
|
0
|
0
|
Rampur
|
0
|
±0.00
|
0
|
±0.00
|
0
|
±0.00
|
0
|
0
|
0
|
Hamirpur
|
Tirasujanpur
|
0
|
±0.00
|
0
|
±0.00
|
0
|
±0.00
|
0
|
0
|
0
|
Nadaun
|
0
|
±0.00
|
0
|
±0.00
|
0
|
±0.00
|
0
|
0
|
0
|
Solan
|
Kandaghat
|
0
|
±0.00
|
0
|
±0.00
|
0
|
±0.00
|
0
|
0
|
0
|
Darlaghat
|
0
|
±0.00
|
0
|
±0.00
|
0
|
±0.00
|
0
|
0
|
0
|
P.D = cyst population density/total no. of samples, % occurence = No. of positive samples / total no. of samples x 100; ± =standard deviation.
Result mentioned in the table.2 explains that the population density and % occurrence of PCN greatly differs from examined locations. The population density and % occurrence of PCN in Distt Mandi were highest and 100%. Whereas Distt Kangra showed the lowest population density and % occurrence. Soil samples collected from Distt Una, Hamirpur, and Solan were showed nil results from particular areas mentioned in the above table. Thus, data recorded during the survey conclude that Distt Mandi had the highest distribution of PCN and lowest in Distt Kangra. Such differences may occur due to location soil type, soil moisture, and irrigation system.
Statistical analysis of the cyst populations reveals some considerable results as given in Table 4 and 5. Table 4 comprehends the summary of P values for the test of differences between a different location within a district and all districts altogether. It can be seen that the cyst population was significantly different between the locations of district Mandi, Chamba, and Kullu. Also, the cyst populations between the districts (all locations altogether) are significantly different (P<0.05). Figure 1-5 depicts the graphical variation of cyst population between the groups.
Pair-wise comparisons between all the groups in concern have been done and its results have been comprehended in table 5. Most of the pairs are not found to be different in the cyst population except a few as mentioned. District Kangra is found to be different from all other districts in the cyst population.
Table 4: with P value for comparison of locations within the district/All districts
Between locations of District
|
P Value
|
Significant? P < 0.05?
|
Test
|
Mandi
|
0.0261
|
Yes
|
Kruskal-Wallis
|
Chamba
|
0.0153
|
Yes
|
Kruskal-Wallis
|
Kullu
|
0.0108
|
Yes
|
Kruskal-Wallis
|
Kangra
|
0.0678
|
No
|
Kruskal-Wallis
|
All districts
|
<0.0001
|
Yes
|
Kruskal-Wallis
|
Table 5: with P value for pair wise comparison of locations/Districts
Locations in consideration
|
Significant? P < 0.05?
|
Mandi District
|
Basunti vs Rewalsar
|
No
|
Basunti vs Phulladhar
|
No
|
Basunti vs Dhanotu
|
No
|
Rewalsar vs Phulladhar
|
No
|
Rewalsar vs Dhanotu
|
No
|
Phulladhar vs Dhanotu
|
Yes
|
Chamba District
|
Saru vs Ahla
|
Yes
|
Saru vs Banikhet
|
No
|
Saru vs Salooni
|
No
|
Ahla vs Banikhet
|
No
|
Ahla vs Salooni
|
No
|
Banikhet vs Salooni
|
No
|
Kullu District
|
Bajaura vs Bandrol
|
No
|
Bajaura vs Anni
|
No
|
Bajaura vs Bhuntar
|
No
|
Bajaura vs Talpura
|
Yes
|
Bandrol vs Anni
|
No
|
Bandrol vs Bhuntar
|
No
|
Bandrol vs Talpura
|
No
|
Anni vs Bhuntar
|
No
|
Anni vs Talpura
|
No
|
Bhuntar vs Talpura
|
No
|
Kangra District
|
Kangra vs Jiya
|
No
|
Kangra vs Skoh
|
No
|
Kangra vs Parihar khar
|
No
|
Kangra vs Shahpur
|
No
|
Kangra vs Malan
|
No
|
Kangra vs Bagoda
|
No
|
Jiya vs Skoh
|
No
|
Jiya vs Parihar khar
|
No
|
Jiya vs Shahpur
|
No
|
Jiya vs Malan
|
No
|
Jiya vs Bagoda
|
No
|
Skoh vs Parihar khar
|
No
|
Skoh vs Shahpur
|
No
|
Skoh vs Malan
|
No
|
Skoh vs Bagoda
|
No
|
Parihar khar vs Shahpur
|
No
|
Parihar khar vs Malan
|
No
|
Parihar khar vs Bagoda
|
No
|
Shahpur vs Malan
|
No
|
Shahpur vs Bagoda
|
No
|
Malan vs Bagoda
|
No
|
All Districts
|
Mandi vs Chamba
|
No
|
Mandi vs Kullu
|
No
|
Mandi vs Kangra
|
Yes
|
Chamba vs Kullu
|
No
|
Chamba vs Kangra
|
Yes
|
Kullu vs Kangra
|
No
|
Globodera spp. was first reported in 1961 from Nilgiris district, Tamil Nadu, India. Ganguli et al. 2010 reported two species G. rostochiensis and G. pallida from the potato breeding farms in Kufri, Shimla District of Himachal Pradesh. District Shimla, Kullu, Mandi, Chamba and Kangra showed the prevalence of PCN. District Shimla, Kullu, Mandi and Kangra showed 100% occurrence of PCN. Chandel et al. 2020 discussed the distribution of PCN in Himachal Pradesh, Jammu and Kashmir and Uttrakhand (northern hilly areas) India. Seenivasan et al. 2017 was conducted study for the distribution of PCN, G. rostochiensis and G. pallida in the potato fields of Kodaikanal hills, Tamil Nadu, India. Showed 36% of potato field is infested with the mixed population of PCN. Due to the increase in the infestation of PCN on potato fields especially in potato breeding farms as results government of India has restricted the supply of potato seeds from the infested areas of these states. There is urgent need of controlling the infestation of PCN on potato. This survey may help in figure out the prevalence and distribution of PCN in Himachal Pradesh and also for planning effective strategies against PCN. However, the use of organophosphorus compounds has been shown to control nematodes but may lead to toxicity and regular use of organophosphorus compounds subside the microbial community and reduces soil fertility (Sidhu et al. 2019). In a study it was observed that even the use of carbendazim can be toxic to soil microorganisms (Singh et al. 2016and that to nematode as well (Whitehead et al. 1985). Camacho et al. 2020 conducted the survey in Portugal during 2013-2019 and reported 28.6% population of G. rostochiensis and 49.4 % population of G. pallida in collected soil samples from different areas. Which finally conclude that G. pallida detected greater than G. rostochiensis in the different potato fields of Portugal. In the way survey of potato field resulted mixed PCN population. But out of two 67% were G. pallida and 8% were G. rostochiensis (Minnis et al. 2002). Infestation of G. pallida is more as compare to G. rostochiensis in England fields. Przetakiewicz 2013 reported the G. rostochiensis pathotypes Ro5 in Poland. G. rostochiensis and G. pallida are also found in island of newfoundland and G. rostochiensis are found in the potato field of Saanich Peninsula of Vancouver Island, British Columbia, Canada (Sun et al. 2007). In Columbia G. pallida is the predominant species in potato cultivating fields and recorded from new areas also. All municipalities area samples were recorded positive (Vallejo et al. 2021). In Ukraine PCN identified on morphological and molecular basis. G. rostochiensis is more prevalent than G. pallida (Pylypenko et al. 2005). As per discussion the prevalence of Globodera spp is very high and there is a urgent need of planning effective and sustainable strategies against PCN.